id sid tid token lemma pos njb-49 1 1 microsoft microsoft PROPN njb-49 1 2 word word NOUN njb-49 1 3 latest late ADJ njb-49 1 4 one.doc one.doc DET njb-49 1 5 nepal nepal ADJ njb-49 1 6     SPACE njb-49 1 7 journal journal NOUN njb-49 1 8     SPACE njb-49 1 9 of of ADP njb-49 1 10     SPACE njb-49 1 11 biotechnology biotechnology NOUN njb-49 1 12 . . PUNCT njb-49 1 13         SPACE njb-49 2 1 jan jan PROPN njb-49 2 2 . . PROPN njb-49 2 3     SPACE njb-49 2 4 2012 2012 NUM njb-49 2 5 , , PUNCT njb-49 2 6     SPACE njb-49 2 7 vol vol NOUN njb-49 2 8 . . PUNCT njb-49 2 9     SPACE njb-49 2 10 2 2 NUM njb-49 2 11 , , PUNCT njb-49 2 12     SPACE njb-49 2 13 no no INTJ njb-49 2 14 . . PUNCT njb-49 2 15     SPACE njb-49 3 1 1 1 NUM njb-49 3 2 : : PUNCT njb-49 3 3     SPACE njb-49 3 4 1 1 X njb-49 3 5     SPACE njb-49 3 6 – – PUNCT njb-49 3 7     SPACE njb-49 3 8 15 15 NUM njb-49 3 9                                                                                                                                                                                                                                         SPACE njb-49 3 10 biotechnology biotechnology NOUN njb-49 3 11     SPACE njb-49 3 12 society society NOUN njb-49 3 13     SPACE njb-49 3 14 of of ADP njb-49 3 15     SPACE njb-49 3 16 nepal nepal NOUN njb-49 3 17     SPACE njb-49 3 18 ( ( PUNCT njb-49 3 19 bsn bsn PROPN njb-49 3 20 ) ) PUNCT njb-49 3 21 , , PUNCT njb-49 3 22     SPACE njb-49 3 23 all all DET njb-49 3 24     SPACE njb-49 3 25 rights right NOUN njb-49 3 26     SPACE njb-49 3 27 reserved reserve VERB njb-49 3 28     SPACE njb-49 3 29 1 1 NUM njb-49 3 30 original original ADJ njb-49 3 31 research research NOUN njb-49 3 32 article article NOUN njb-49 3 33 cloning cloning NOUN njb-49 3 34 , , PUNCT njb-49 3 35     SPACE njb-49 3 36 expression expression NOUN njb-49 3 37 , , PUNCT njb-49 3 38     SPACE njb-49 3 39 purification purification NOUN njb-49 3 40 , , PUNCT njb-49 3 41     SPACE njb-49 3 42 and and CCONJ njb-49 3 43     SPACE njb-49 3 44 characterization characterization NOUN njb-49 3 45     SPACE njb-49 3 46 of of ADP njb-49 3 47     SPACE njb-49 3 48 clostridium clostridium NOUN njb-49 3 49     SPACE njb-49 3 50 botulinum botulinum ADJ njb-49 3 51     SPACE njb-49 3 52 neurotoxin neurotoxin NOUN njb-49 3 53     SPACE njb-49 3 54 serotype serotype NOUN njb-49 3 55     SPACE njb-49 3 56 f f PROPN njb-49 3 57     SPACE njb-49 3 58 domains domain NOUN njb-49 3 59     SPACE njb-49 3 60 santosh santosh PROPN njb-49 3 61 dulal1 dulal1 PROPN njb-49 3 62 * * PROPN njb-49 3 63 , , PUNCT njb-49 3 64 bhupal bhupal PROPN njb-49 3 65 ban2 ban2 PROPN njb-49 3 66 , , PUNCT njb-49 3 67 gi gi PROPN njb-49 3 68 hyoek hyoek X njb-49 3 69 yang2 yang2 PROPN njb-49 3 70 , , PUNCT njb-49 3 71 and and CCONJ njb-49 3 72 hyun hyun PROPN njb-49 3 73 ho ho PROPN njb-49 3 74 jung1 jung1 PROPN njb-49 3 75 , , PUNCT njb-49 3 76 2 2 NUM njb-49 3 77 1department 1department NUM njb-49 3 78 of of ADP njb-49 3 79 applied apply VERB njb-49 3 80 molecular molecular ADJ njb-49 3 81 biology biology NOUN njb-49 3 82 , , PUNCT njb-49 3 83 sun sun PROPN njb-49 3 84 moon moon PROPN njb-49 3 85 university university PROPN njb-49 3 86 , , PUNCT njb-49 3 87 asan asan PROPN njb-49 3 88 - - PUNCT njb-49 3 89 si si PROPN njb-49 3 90 , , PUNCT njb-49 3 91 chung chung PROPN njb-49 3 92 - - PUNCT njb-49 3 93 nam nam PROPN njb-49 3 94 , , PUNCT njb-49 3 95 336 336 NUM njb-49 3 96 - - SYM njb-49 3 97 708 708 NUM njb-49 3 98 , , PUNCT njb-49 3 99 the the DET njb-49 3 100 republic republic PROPN njb-49 3 101 of of ADP njb-49 3 102 korea korea PROPN njb-49 3 103 2medy 2medy PROPN njb-49 3 104 - - PUNCT njb-49 3 105 tox tox PROPN njb-49 3 106 inc inc PROPN njb-49 3 107 . . PROPN njb-49 3 108 , , PUNCT njb-49 3 109 cheongwon cheongwon PROPN njb-49 3 110 - - PUNCT njb-49 3 111 gun gun NOUN njb-49 3 112 , , PUNCT njb-49 3 113 chungbuk chungbuk PROPN njb-49 3 114 - - PUNCT njb-49 3 115 do do VERB njb-49 3 116 , , PUNCT njb-49 3 117 363 363 NUM njb-49 3 118 - - SYM njb-49 3 119 883 883 NUM njb-49 3 120 , , PUNCT njb-49 3 121 the the DET njb-49 3 122 republic republic PROPN njb-49 3 123 of of ADP njb-49 3 124 korea korea PROPN njb-49 3 125 * * PUNCT njb-49 3 126 correspondence correspondence NOUN njb-49 3 127 author author NOUN njb-49 3 128 recent recent ADJ njb-49 3 129 address address NOUN njb-49 3 130 : : PUNCT njb-49 3 131 microbiology microbiology NOUN njb-49 3 132 graduate graduate NOUN njb-49 3 133 program program NOUN njb-49 3 134 , , PUNCT njb-49 3 135 department department NOUN njb-49 3 136 of of ADP njb-49 3 137 biology biology NOUN njb-49 3 138 , , PUNCT njb-49 3 139 new new PROPN njb-49 3 140 mexico mexico PROPN njb-49 3 141 state state PROPN njb-49 3 142 university university PROPN njb-49 3 143 , , PUNCT njb-49 3 144 las las PROPN njb-49 3 145 cruces crux NOUN njb-49 3 146 , , PUNCT njb-49 3 147 nm nm PROPN njb-49 3 148 88003 88003 NUM njb-49 3 149 , , PUNCT njb-49 3 150 united united PROPN njb-49 3 151 states states PROPN njb-49 3 152 of of ADP njb-49 3 153 america america PROPN njb-49 3 154 . . PUNCT njb-49 4 1 e e X njb-49 4 2 - - NOUN njb-49 4 3 mail mail NOUN njb-49 4 4 : : PUNCT njb-49 4 5 santoshd@nmsu.edu santoshd@nmsu.edu NOUN njb-49 4 6 abstract abstract ADJ njb-49 4 7 the the DET njb-49 4 8 use use NOUN njb-49 4 9 of of ADP njb-49 4 10 recombinant recombinant ADJ njb-49 4 11 bont bont ADJ njb-49 4 12 domains domain NOUN njb-49 4 13 has have AUX njb-49 4 14 been be AUX njb-49 4 15 proposed propose VERB njb-49 4 16 as as ADP njb-49 4 17 a a DET njb-49 4 18 means means NOUN njb-49 4 19 to to PART njb-49 4 20 develop develop VERB njb-49 4 21 strategies strategy NOUN njb-49 4 22 to to PART njb-49 4 23 treat treat VERB njb-49 4 24 and and CCONJ njb-49 4 25 prevent prevent VERB njb-49 4 26 botulism botulism NOUN njb-49 4 27 . . PUNCT njb-49 5 1 here here ADV njb-49 5 2 , , PUNCT njb-49 5 3 details detail NOUN njb-49 5 4 on on ADP njb-49 5 5 the the DET njb-49 5 6 molecular molecular ADJ njb-49 5 7 cloning cloning NOUN njb-49 5 8 , , PUNCT njb-49 5 9 protein protein NOUN njb-49 5 10 expression expression NOUN njb-49 5 11 , , PUNCT njb-49 5 12 purification purification NOUN njb-49 5 13 , , PUNCT njb-49 5 14 and and CCONJ njb-49 5 15 immunoreactivity immunoreactivity NOUN njb-49 5 16 of of ADP njb-49 5 17 bont bont PROPN njb-49 5 18 / / SYM njb-49 5 19 f f PROPN njb-49 5 20 domains domain NOUN njb-49 5 21 from from ADP njb-49 5 22 clostridium clostridium NOUN njb-49 5 23 botulinum botulinum NOUN njb-49 5 24 are be AUX njb-49 5 25 presented present VERB njb-49 5 26 . . PUNCT njb-49 6 1 initially initially ADV njb-49 6 2 , , PUNCT njb-49 6 3 fulllength fulllength PROPN njb-49 6 4 synthetic synthetic ADJ njb-49 6 5 genes gene NOUN njb-49 6 6 encoding encode VERB njb-49 6 7 recombinant recombinant ADJ njb-49 6 8 bont bont NOUN njb-49 6 9 / / SYM njb-49 6 10 f f PROPN njb-49 6 11 domains domain NOUN njb-49 6 12 ( ( PUNCT njb-49 6 13 catalytic catalytic ADJ njb-49 6 14 , , PUNCT njb-49 6 15 translocation translocation NOUN njb-49 6 16 , , PUNCT njb-49 6 17 and and CCONJ njb-49 6 18 receptor receptor NOUN njb-49 6 19 binding binding NOUN njb-49 6 20 ) ) PUNCT njb-49 6 21 were be AUX njb-49 6 22 designed design VERB njb-49 6 23 and and CCONJ njb-49 6 24 cloned clone VERB njb-49 6 25 into into ADP njb-49 6 26 escherichia escherichia NOUN njb-49 6 27 coli coli NOUN njb-49 6 28 for for ADP njb-49 6 29 expression expression NOUN njb-49 6 30 . . PUNCT njb-49 7 1 recombinant recombinant ADJ njb-49 7 2 proteins protein NOUN njb-49 7 3 were be AUX njb-49 7 4 then then ADV njb-49 7 5 purified purify VERB njb-49 7 6 through through ADP njb-49 7 7 gst gst PROPN njb-49 7 8 affinity affinity NOUN njb-49 7 9 chromatography chromatography NOUN njb-49 7 10 preceding precede VERB njb-49 7 11 elution elution NOUN njb-49 7 12 of of ADP njb-49 7 13 gst gst NOUN njb-49 7 14 - - PUNCT njb-49 7 15 free free ADJ njb-49 7 16 recombinant recombinant ADJ njb-49 7 17 domains domain NOUN njb-49 7 18 by by ADP njb-49 7 19 thrombin thrombin NOUN njb-49 7 20 protease protease NOUN njb-49 7 21 . . PUNCT njb-49 8 1 soluble soluble ADJ njb-49 8 2 recombinant recombinant ADJ njb-49 8 3 proteins protein NOUN njb-49 8 4 encoding encode VERB njb-49 8 5 catalytic catalytic ADJ njb-49 8 6 light light NOUN njb-49 8 7 chain chain NOUN njb-49 8 8 and and CCONJ njb-49 8 9 translocation translocation NOUN njb-49 8 10 n n ADJ njb-49 8 11 - - ADJ njb-49 8 12 terminal terminal ADJ njb-49 8 13 heavy heavy ADJ njb-49 8 14 chain chain NOUN njb-49 8 15 were be AUX njb-49 8 16 subsequently subsequently ADV njb-49 8 17 used use VERB njb-49 8 18 to to PART njb-49 8 19 perform perform VERB njb-49 8 20 in in ADP njb-49 8 21 vivo vivo ADJ njb-49 8 22 immunization immunization NOUN njb-49 8 23 . . PUNCT njb-49 9 1 polyclonal polyclonal ADJ njb-49 9 2 mouse mouse NOUN njb-49 9 3 antibodies antibodie VERB njb-49 9 4 specific specific ADJ njb-49 9 5 to to ADP njb-49 9 6 these these DET njb-49 9 7 domains domain NOUN njb-49 9 8 were be AUX njb-49 9 9 raised raise VERB njb-49 9 10 , , PUNCT njb-49 9 11 confirmed confirm VERB njb-49 9 12 by by ADP njb-49 9 13 western western ADJ njb-49 9 14 blot blot NOUN njb-49 9 15 analysis analysis NOUN njb-49 9 16 and and CCONJ njb-49 9 17 elevated elevated ADJ njb-49 9 18 immunoreactivity immunoreactivity NOUN njb-49 9 19 was be AUX njb-49 9 20 identified identify VERB njb-49 9 21 through through ADP njb-49 9 22 indirect indirect ADJ njb-49 9 23 elisa elisa NOUN njb-49 9 24 . . PUNCT njb-49 10 1 in in ADP njb-49 10 2 conclusion conclusion NOUN njb-49 10 3 , , PUNCT njb-49 10 4 availability availability NOUN njb-49 10 5 of of ADP njb-49 10 6 the the DET njb-49 10 7 recombinant recombinant ADJ njb-49 10 8 protein protein NOUN njb-49 10 9 provides provide VERB njb-49 10 10 an an DET njb-49 10 11 effective effective ADJ njb-49 10 12 system system NOUN njb-49 10 13 to to PART njb-49 10 14 study study VERB njb-49 10 15 the the DET njb-49 10 16 immunological immunological ADJ njb-49 10 17 aspects aspect NOUN njb-49 10 18 of of ADP njb-49 10 19 bont bont PROPN njb-49 10 20 / / SYM njb-49 10 21 f f PROPN njb-49 10 22 and and CCONJ njb-49 10 23 corresponding corresponding ADJ njb-49 10 24 applications application NOUN njb-49 10 25 in in ADP njb-49 10 26 pathogen pathogen PROPN njb-49 10 27 detection detection NOUN njb-49 10 28 and and CCONJ njb-49 10 29 vaccine vaccine NOUN njb-49 10 30 candidacy candidacy NOUN njb-49 10 31 . . PUNCT njb-49 11 1 keywords keyword NOUN njb-49 11 2 : : PUNCT njb-49 11 3 clostridium clostridium NOUN njb-49 11 4 botulunium botulunium NOUN njb-49 11 5 ; ; PUNCT njb-49 11 6 botulinum botulinum ADJ njb-49 11 7 neurotoxin neurotoxin NOUN njb-49 11 8 type type NOUN njb-49 11 9 f f PROPN njb-49 11 10 ( ( PUNCT njb-49 11 11 bont bont PROPN njb-49 11 12 / / SYM njb-49 11 13 f f PROPN njb-49 11 14 ) ) PUNCT njb-49 11 15 domains domain NOUN njb-49 11 16 ; ; PUNCT njb-49 11 17 cloning cloning NOUN njb-49 11 18 ; ; PUNCT njb-49 11 19 recombinant recombinant ADJ njb-49 11 20 protein protein NOUN njb-49 11 21 expression expression NOUN njb-49 11 22 ; ; PUNCT njb-49 11 23 immunoreactivity immunoreactivity NOUN njb-49 11 24 introduction introduction NOUN njb-49 11 25 botulinum botulinum NOUN njb-49 11 26 neurotoxins neurotoxin NOUN njb-49 11 27 ( ( PUNCT njb-49 11 28 bonts bont NOUN njb-49 11 29 ) ) PUNCT njb-49 11 30 , , PUNCT njb-49 11 31 the the DET njb-49 11 32 most most ADV njb-49 11 33 potent potent ADJ njb-49 11 34 of of ADP njb-49 11 35 all all DET njb-49 11 36 biological biological ADJ njb-49 11 37 substances substance NOUN njb-49 11 38 known know VERB njb-49 11 39 to to ADP njb-49 11 40 date date NOUN njb-49 11 41 , , PUNCT njb-49 11 42 are be AUX njb-49 11 43 produced produce VERB njb-49 11 44 by by ADP njb-49 11 45 several several ADJ njb-49 11 46 species specie NOUN njb-49 11 47 of of ADP njb-49 11 48 the the DET njb-49 11 49 genera genera NOUN njb-49 11 50 clostridia clostridia PROPN njb-49 11 51 ( ( PUNCT njb-49 11 52 c. c. PROPN njb-49 11 53 botulinum botulinum PROPN njb-49 11 54 , , PUNCT njb-49 11 55 c. c. PROPN njb-49 11 56 butyricum butyricum PROPN njb-49 11 57 , , PUNCT njb-49 11 58 and and CCONJ njb-49 11 59 c. c. PROPN njb-49 11 60 baratii baratii PROPN njb-49 11 61 ) ) PUNCT njb-49 12 1 [ [ X njb-49 12 2 1 1 NUM njb-49 12 3 - - SYM njb-49 12 4 3 3 NUM njb-49 12 5 ] ] PUNCT njb-49 12 6 . . PUNCT njb-49 13 1 seven seven NUM njb-49 13 2 immunologically immunologically ADV njb-49 13 3 distinct distinct ADJ njb-49 13 4 botulinum botulinum ADJ njb-49 13 5 neurotoxin neurotoxin NOUN njb-49 13 6 serotypes serotype VERB njb-49 13 7 bonts bont NOUN njb-49 13 8 / / SYM njb-49 13 9 a a DET njb-49 13 10 - - PUNCT njb-49 13 11 g g NOUN njb-49 13 12 are be AUX njb-49 13 13 produced produce VERB njb-49 13 14 and and CCONJ njb-49 13 15 implicated implicate VERB njb-49 13 16 in in ADP njb-49 13 17 botulism botulism NOUN njb-49 13 18 poisoning poison VERB njb-49 13 19 [ [ X njb-49 13 20 4 4 NUM njb-49 13 21 ] ] X njb-49 13 22 . . PUNCT njb-49 14 1 botulism botulism NOUN njb-49 14 2 is be AUX njb-49 14 3 a a DET njb-49 14 4 serious serious ADJ njb-49 14 5 neuroparalytic neuroparalytic ADJ njb-49 14 6 disease disease NOUN njb-49 14 7 [ [ X njb-49 14 8 5 5 NUM njb-49 14 9 ] ] X njb-49 14 10 , , PUNCT njb-49 14 11 which which PRON njb-49 14 12 generally generally ADV njb-49 14 13 occurs occur VERB njb-49 14 14 through through ADP njb-49 14 15 ingestion ingestion NOUN njb-49 14 16 of of ADP njb-49 14 17 preformed preform VERB njb-49 14 18 toxin toxin NOUN njb-49 14 19 or or CCONJ njb-49 14 20 rarely rarely ADV njb-49 14 21 , , PUNCT njb-49 14 22 through through ADP njb-49 14 23 infection infection NOUN njb-49 14 24 of of ADP njb-49 14 25 wounds wound NOUN njb-49 14 26 . . PUNCT njb-49 15 1 the the DET njb-49 15 2 center center NOUN njb-49 15 3 for for ADP njb-49 15 4 disease disease NOUN njb-49 15 5 control control NOUN njb-49 15 6 and and CCONJ njb-49 15 7 prevention prevention NOUN njb-49 15 8 ( ( PUNCT njb-49 15 9 cdc cdc PROPN njb-49 15 10 ) ) PUNCT njb-49 15 11 classifies classify VERB njb-49 15 12 bonts bont NOUN njb-49 15 13 , , PUNCT njb-49 15 14 among among ADP njb-49 15 15 the the DET njb-49 15 16 six six NUM njb-49 15 17 highest high ADJ njb-49 15 18 risk risk NOUN njb-49 15 19 threat threat NOUN njb-49 15 20 agents agent NOUN njb-49 15 21 for for ADP njb-49 15 22 bioterrorism bioterrorism NOUN njb-49 15 23 “ " PUNCT njb-49 15 24 class class NOUN njb-49 15 25 a a DET njb-49 15 26 biological biological ADJ njb-49 15 27 warfare warfare NOUN njb-49 15 28 agent agent NOUN njb-49 15 29 ” " PUNCT njb-49 16 1 [ [ PUNCT njb-49 16 2 6 6 NUM njb-49 16 3 , , PUNCT njb-49 16 4 7 7 NUM njb-49 16 5 ] ] PUNCT njb-49 16 6 . . PUNCT njb-49 17 1 despite despite SCONJ njb-49 17 2 their their PRON njb-49 17 3 potential potential NOUN njb-49 17 4 to to PART njb-49 17 5 be be AUX njb-49 17 6 used use VERB njb-49 17 7 for for ADP njb-49 17 8 deleterious deleterious ADJ njb-49 17 9 purposes purpose NOUN njb-49 17 10 , , PUNCT njb-49 17 11 bonts bont NOUN njb-49 17 12 have have AUX njb-49 17 13 increasing increase VERB njb-49 17 14 nepal nepal ADJ njb-49 17 15     SPACE njb-49 17 16 journal journal NOUN njb-49 17 17     SPACE njb-49 17 18 of of ADP njb-49 17 19     SPACE njb-49 17 20 biotechnology biotechnology NOUN njb-49 17 21 . . PUNCT njb-49 17 22         SPACE njb-49 18 1 jan jan PROPN njb-49 18 2 . . PROPN njb-49 18 3     SPACE njb-49 18 4 2012 2012 NUM njb-49 18 5 , , PUNCT njb-49 18 6     SPACE njb-49 18 7 vol vol NOUN njb-49 18 8 . . PUNCT njb-49 18 9     SPACE njb-49 18 10 2 2 NUM njb-49 18 11 , , PUNCT njb-49 18 12     SPACE njb-49 18 13 no no INTJ njb-49 18 14 . . PUNCT njb-49 18 15     SPACE njb-49 19 1 1 1 NUM njb-49 19 2 : : PUNCT njb-49 19 3     SPACE njb-49 19 4 1 1 X njb-49 19 5     SPACE njb-49 19 6 – – PUNCT njb-49 19 7     SPACE njb-49 19 8 15 15 NUM njb-49 19 9                                                                                                                                                                                                                                         SPACE njb-49 19 10 biotechnology biotechnology NOUN njb-49 19 11     SPACE njb-49 19 12 society society NOUN njb-49 19 13     SPACE njb-49 19 14 of of ADP njb-49 19 15     SPACE njb-49 19 16 nepal nepal NOUN njb-49 19 17     SPACE njb-49 19 18 ( ( PUNCT njb-49 19 19 bsn bsn PROPN njb-49 19 20 ) ) PUNCT njb-49 19 21 , , PUNCT njb-49 19 22     SPACE njb-49 19 23 all all DET njb-49 19 24     SPACE njb-49 19 25 rights right NOUN njb-49 19 26     SPACE njb-49 19 27 reserved reserve VERB njb-49 19 28     SPACE njb-49 19 29 2 2 NUM njb-49 19 30 applications application NOUN njb-49 19 31 in in ADP njb-49 19 32 cosmetics cosmetic NOUN njb-49 19 33 [ [ PUNCT njb-49 19 34 8 8 NUM njb-49 19 35 ] ] PUNCT njb-49 19 36 and and CCONJ njb-49 19 37 therapeutics therapeutic NOUN njb-49 19 38 for for ADP njb-49 19 39 the the DET njb-49 19 40 treatment treatment NOUN njb-49 19 41 of of ADP njb-49 19 42 numerous numerous ADJ njb-49 19 43 dystonias dystonia NOUN njb-49 19 44 , , PUNCT njb-49 19 45 inflammation inflammation NOUN njb-49 19 46 , , PUNCT njb-49 19 47 and and CCONJ njb-49 19 48 chronic chronic ADJ njb-49 19 49 pain pain NOUN njb-49 19 50 [ [ PUNCT njb-49 19 51 9 9 NUM njb-49 19 52 - - SYM njb-49 19 53 11 11 NUM njb-49 19 54 ] ] PUNCT njb-49 19 55 . . PUNCT njb-49 20 1 botulinum botulinum PROPN njb-49 20 2 neurotoxin neurotoxin NOUN njb-49 20 3 serotype serotype PROPN njb-49 20 4 f f PROPN njb-49 21 1 ( ( PUNCT njb-49 21 2 bont bont PROPN njb-49 21 3 / / SYM njb-49 21 4 f f PROPN njb-49 21 5 ) ) PUNCT njb-49 21 6 , , PUNCT njb-49 21 7 is be AUX njb-49 21 8 a a DET njb-49 21 9 member member NOUN njb-49 21 10 of of ADP njb-49 21 11 the the DET njb-49 21 12 botulinum botulinum ADJ njb-49 21 13 neurotoxin neurotoxin NOUN njb-49 21 14 family family NOUN njb-49 21 15 as as ADP njb-49 21 16 a a DET njb-49 21 17 single single ADJ njb-49 21 18 ~150 ~150 NUM njb-49 21 19 kda kda NOUN njb-49 21 20 inactive inactive ADJ njb-49 21 21 polypeptide polypeptide NOUN njb-49 21 22 chain chain NOUN njb-49 21 23 post post ADJ njb-49 21 24 - - ADJ njb-49 21 25 translationally translationally ADV njb-49 21 26 nicked nick VERB njb-49 21 27 , , PUNCT njb-49 21 28 forming form VERB njb-49 21 29 a a DET njb-49 21 30 dichain dichain NOUN njb-49 21 31 consisting consist VERB njb-49 21 32 of of ADP njb-49 21 33 a a DET njb-49 21 34 c c PROPN njb-49 21 35 - - PUNCT njb-49 21 36 terminal terminal ADJ njb-49 21 37 ~100 ~100 NUM njb-49 21 38 kda kda NOUN njb-49 21 39 heavy heavy ADJ njb-49 21 40 chain chain NOUN njb-49 21 41 ( ( PUNCT njb-49 21 42 hc hc INTJ njb-49 21 43 ) ) PUNCT njb-49 21 44 and and CCONJ njb-49 21 45 a a DET njb-49 21 46 n n NOUN njb-49 21 47 - - PUNCT njb-49 21 48 terminal terminal ADJ njb-49 21 49 ~50 ~50 NUM njb-49 21 50 kda kda NOUN njb-49 21 51 light light NOUN njb-49 21 52 chain chain NOUN njb-49 21 53 ( ( PUNCT njb-49 21 54 lc lc NOUN njb-49 21 55 ) ) PUNCT njb-49 21 56 linked link VERB njb-49 21 57 by by ADP njb-49 21 58 a a DET njb-49 21 59 disulphide disulphide ADJ njb-49 21 60 bond bond NOUN njb-49 21 61 [ [ PUNCT njb-49 21 62 12 12 NUM njb-49 21 63 , , PUNCT njb-49 21 64 13 13 NUM njb-49 21 65 ] ] PUNCT njb-49 21 66 . . PUNCT njb-49 22 1 bont bont PROPN njb-49 22 2 / / SYM njb-49 22 3 f f PROPN njb-49 22 4 cleaves cleave VERB njb-49 22 5 its its PRON njb-49 22 6 substrate substrate ADJ njb-49 22 7 vesicle vesicle NOUN njb-49 22 8 associated associate VERB njb-49 22 9 membrane membrane NOUN njb-49 22 10 protein protein NOUN njb-49 22 11 ( ( PUNCT njb-49 22 12 vamp vamp NOUN njb-49 22 13 ) ) PUNCT njb-49 22 14 at at ADP njb-49 22 15 position position NOUN njb-49 22 16 ( ( PUNCT njb-49 22 17 gln58 gln58 NOUN njb-49 22 18 - - PUNCT njb-49 22 19 lys59 lys59 ADJ njb-49 22 20 ) ) PUNCT njb-49 22 21 [ [ PUNCT njb-49 22 22 14 14 NUM njb-49 22 23 , , PUNCT njb-49 22 24 15 15 NUM njb-49 22 25 ] ] PUNCT njb-49 22 26 , , PUNCT njb-49 22 27 one one NUM njb-49 22 28 of of ADP njb-49 22 29 three three NUM njb-49 22 30 neuronal neuronal ADJ njb-49 22 31 proteins protein NOUN njb-49 22 32 associated associate VERB njb-49 22 33 with with ADP njb-49 22 34 exocytosis exocytosis NOUN njb-49 22 35 ; ; PUNCT njb-49 22 36 subsequently subsequently ADV njb-49 22 37 inhibiting inhibit VERB njb-49 22 38 acetylcholine acetylcholine NOUN njb-49 22 39 release release NOUN njb-49 22 40 , , PUNCT njb-49 22 41 resulting result VERB njb-49 22 42 in in ADP njb-49 22 43 death death NOUN njb-49 22 44 by by ADP njb-49 22 45 flaccid flaccid ADJ njb-49 22 46 paralysis paralysis NOUN njb-49 22 47 [ [ PUNCT njb-49 22 48 16 16 NUM njb-49 22 49 , , PUNCT njb-49 22 50 17 17 NUM njb-49 22 51 ] ] PUNCT njb-49 22 52 . . PUNCT njb-49 23 1 each each DET njb-49 23 2 bont bont PROPN njb-49 23 3 / / SYM njb-49 23 4 f f PROPN njb-49 23 5 partial partial ADJ njb-49 23 6 fragment fragment NOUN njb-49 23 7 ; ; PUNCT njb-49 23 8 light light ADJ njb-49 23 9 chain chain NOUN njb-49 23 10 catalytic catalytic ADJ njb-49 23 11 domain domain NOUN njb-49 23 12 ( ( PUNCT njb-49 23 13 rf rf NOUN njb-49 23 14 - - PUNCT njb-49 23 15 lc lc NOUN njb-49 23 16 ) ) PUNCT njb-49 23 17 , , PUNCT njb-49 23 18 n n NOUN njb-49 23 19 - - ADJ njb-49 23 20 terminal terminal ADJ njb-49 23 21 half half NOUN njb-49 23 22 of of ADP njb-49 23 23 the the DET njb-49 23 24 heavy heavy ADJ njb-49 23 25 chain chain NOUN njb-49 23 26 translocation translocation NOUN njb-49 23 27 domain domain NOUN njb-49 23 28 ( ( PUNCT njb-49 23 29 rfhn rfhn PROPN njb-49 23 30 ) ) PUNCT njb-49 23 31 , , PUNCT njb-49 23 32 the the DET njb-49 23 33 c c X njb-49 23 34 - - PUNCT njb-49 23 35 terminal terminal ADJ njb-49 23 36 half half NOUN njb-49 23 37 of of ADP njb-49 23 38 the the DET njb-49 23 39 heavy heavy ADJ njb-49 23 40 chain chain NOUN njb-49 23 41 receptor receptor NOUN njb-49 23 42 binding bind VERB njb-49 23 43 domain domain NOUN njb-49 23 44 ( ( PUNCT njb-49 23 45 rf rf NOUN njb-49 23 46 - - PUNCT njb-49 23 47 hc hc NOUN njb-49 23 48 ) ) PUNCT njb-49 23 49 , , PUNCT njb-49 23 50 and and CCONJ njb-49 23 51 cterminal cterminal ADJ njb-49 23 52 quarter quarter NOUN njb-49 23 53 part part NOUN njb-49 23 54 of of ADP njb-49 23 55 the the DET njb-49 23 56 heavy heavy ADJ njb-49 23 57 chain chain NOUN njb-49 23 58 receptor receptor NOUN njb-49 23 59 binding bind VERB njb-49 23 60 domain domain NOUN njb-49 23 61 ( ( PUNCT njb-49 23 62 rf rf NOUN njb-49 23 63 - - PUNCT njb-49 23 64 hcc hcc PROPN njb-49 23 65 ) ) PUNCT njb-49 23 66 plays play VERB njb-49 23 67 a a DET njb-49 23 68 specific specific ADJ njb-49 23 69 role role NOUN njb-49 23 70 in in ADP njb-49 23 71 the the DET njb-49 23 72 toxicity toxicity NOUN njb-49 23 73 mechanism mechanism NOUN njb-49 23 74 [ [ PUNCT njb-49 23 75 18 18 NUM njb-49 23 76 , , PUNCT njb-49 23 77 19 19 NUM njb-49 23 78 ] ] PUNCT njb-49 23 79 . . PUNCT njb-49 24 1 botulinum botulinum PROPN njb-49 24 2 neurotoxin neurotoxin PROPN njb-49 24 3 lcs lcs PROPN njb-49 24 4 operate operate VERB njb-49 24 5 by by ADP njb-49 24 6 zinc zinc NOUN njb-49 24 7 dependent dependent ADJ njb-49 24 8 proteolysis proteolysis NOUN njb-49 24 9 involved involve VERB njb-49 24 10 in in ADP njb-49 24 11 neurotransmitter neurotransmitter NOUN njb-49 24 12 exocytosis exocytosis NOUN njb-49 24 13 from from ADP njb-49 24 14 presynaptic presynaptic ADJ njb-49 24 15 termini termini NOUN njb-49 25 1 [ [ X njb-49 25 2 20 20 NUM njb-49 25 3 ] ] PUNCT njb-49 25 4 . . PUNCT njb-49 26 1 botulinum botulinum PROPN njb-49 26 2 neurotoxin neurotoxin PROPN njb-49 26 3 hns hns PROPN njb-49 26 4 possesses possess VERB njb-49 26 5 channel channel NOUN njb-49 26 6 - - PUNCT njb-49 26 7 forming form VERB njb-49 26 8 capability capability NOUN njb-49 26 9 in in ADP njb-49 26 10 the the DET njb-49 26 11 acidic acidic ADJ njb-49 26 12 environment environment NOUN njb-49 26 13 of of ADP njb-49 26 14 the the DET njb-49 26 15 endosome endosome NOUN njb-49 26 16 , , PUNCT njb-49 26 17 allowing allow VERB njb-49 26 18 internalization internalization NOUN njb-49 26 19 of of ADP njb-49 26 20 the the DET njb-49 26 21 toxin toxin NOUN njb-49 26 22 , , PUNCT njb-49 26 23 while while SCONJ njb-49 26 24 hcs hcs NOUN njb-49 26 25 are be AUX njb-49 26 26 involved involve VERB njb-49 26 27 in in ADP njb-49 26 28 specific specific ADJ njb-49 26 29 binding binding NOUN njb-49 26 30 to to ADP njb-49 26 31 the the DET njb-49 26 32 presynaptic presynaptic ADJ njb-49 26 33 membrane membrane NOUN njb-49 26 34 via via ADP njb-49 26 35 gangliosides ganglioside NOUN njb-49 26 36 and and CCONJ njb-49 26 37 a a DET njb-49 26 38 protein protein NOUN njb-49 26 39 coreceptor coreceptor NOUN njb-49 26 40 [ [ PUNCT njb-49 26 41 21 21 NUM njb-49 26 42 ] ] PUNCT njb-49 26 43 . . PUNCT njb-49 27 1 lastly lastly ADV njb-49 27 2 , , PUNCT njb-49 27 3 the the DET njb-49 27 4 hcc hcc PROPN njb-49 27 5 region region PROPN njb-49 27 6 of of ADP njb-49 27 7 bonts bont NOUN njb-49 27 8 are be AUX njb-49 27 9 known know VERB njb-49 27 10 to to PART njb-49 27 11 harbor harbor VERB njb-49 27 12 the the DET njb-49 27 13 receptor receptor NOUN njb-49 27 14 binding bind VERB njb-49 27 15 neutralizing neutralizing ADJ njb-49 27 16 epitopes epitope NOUN njb-49 27 17 which which PRON njb-49 27 18 are be AUX njb-49 27 19 targets target NOUN njb-49 27 20 for for ADP njb-49 27 21 antibodies antibody NOUN njb-49 27 22 that that PRON njb-49 27 23 can can AUX njb-49 27 24 specifically specifically ADV njb-49 27 25 bind bind VERB njb-49 27 26 to to ADP njb-49 27 27 the the DET njb-49 27 28 receptor receptor NOUN njb-49 27 29 , , PUNCT njb-49 27 30 and and CCONJ njb-49 27 31 show show VERB njb-49 27 32 neutralizing neutralize VERB njb-49 27 33 activity activity NOUN njb-49 27 34 against against ADP njb-49 27 35 bont bont ADJ njb-49 27 36 toxicity toxicity NOUN njb-49 27 37 [ [ PUNCT njb-49 27 38 22 22 NUM njb-49 27 39 ] ] PUNCT njb-49 27 40 . . PUNCT njb-49 28 1 thus thus ADV njb-49 28 2 , , PUNCT njb-49 28 3 it it PRON njb-49 28 4 is be AUX njb-49 28 5 imperative imperative ADJ njb-49 28 6 to to PART njb-49 28 7 construct construct VERB njb-49 28 8 each each DET njb-49 28 9 domain domain NOUN njb-49 28 10 for for ADP njb-49 28 11 the the DET njb-49 28 12 analysis analysis NOUN njb-49 28 13 of of ADP njb-49 28 14 their their PRON njb-49 28 15 molecular molecular ADJ njb-49 28 16 and and CCONJ njb-49 28 17 biochemical biochemical ADJ njb-49 28 18 activities activity NOUN njb-49 28 19 as as ADV njb-49 28 20 well well ADV njb-49 28 21 as as ADP njb-49 28 22 for for ADP njb-49 28 23 the the DET njb-49 28 24 development development NOUN njb-49 28 25 of of ADP njb-49 28 26 potential potential ADJ njb-49 28 27 neutralizing neutralizing ADJ njb-49 28 28 antibodies antibody NOUN njb-49 28 29 specific specific ADJ njb-49 28 30 to to ADP njb-49 28 31 each each PRON njb-49 28 32 . . PUNCT njb-49 29 1 to to PART njb-49 29 2 achieve achieve VERB njb-49 29 3 this this DET njb-49 29 4 goal goal NOUN njb-49 29 5 , , PUNCT njb-49 29 6 bont bont PROPN njb-49 29 7 / / SYM njb-49 29 8 f f PROPN njb-49 29 9 domains domain NOUN njb-49 29 10 from from ADP njb-49 29 11 c. c. PROPN njb-49 29 12 botulinum botulinum PROPN njb-49 29 13 were be AUX njb-49 29 14 cloned clone VERB njb-49 29 15 and and CCONJ njb-49 29 16 expressed express VERB njb-49 29 17 using use VERB njb-49 29 18 a a DET njb-49 29 19 high high ADJ njb-49 29 20 expression expression NOUN njb-49 29 21 vector vector NOUN njb-49 29 22 and and CCONJ njb-49 29 23 compatible compatible ADJ njb-49 29 24 host host NOUN njb-49 29 25 e. e. PROPN njb-49 29 26 coli coli PROPN njb-49 29 27 strains strain VERB njb-49 29 28 to to PART njb-49 29 29 obtain obtain VERB njb-49 29 30 high high ADJ njb-49 29 31 quality quality NOUN njb-49 29 32 of of ADP njb-49 29 33 recombinant recombinant ADJ njb-49 29 34 proteins protein NOUN njb-49 29 35 suitable suitable ADJ njb-49 29 36 for for ADP njb-49 29 37 administration administration NOUN njb-49 29 38 into into ADP njb-49 29 39 mice mouse NOUN njb-49 29 40 . . PUNCT njb-49 30 1 methods method NOUN njb-49 30 2 chemicals chemical NOUN njb-49 30 3 , , PUNCT njb-49 30 4 buffers buffer NOUN njb-49 30 5 , , PUNCT njb-49 30 6 and and CCONJ njb-49 30 7 reagents reagent NOUN njb-49 30 8 components component NOUN njb-49 30 9 related relate VERB njb-49 30 10 to to ADP njb-49 30 11 dna dna PROPN njb-49 30 12 manipulation manipulation NOUN njb-49 30 13 , , PUNCT njb-49 30 14 including include VERB njb-49 30 15 ex ex NOUN njb-49 30 16 - - ADJ njb-49 30 17 taq taq ADJ njb-49 30 18 polymerase polymerase NOUN njb-49 30 19 , , PUNCT njb-49 30 20 dntp dntp NOUN njb-49 30 21 , , PUNCT njb-49 30 22 and and CCONJ njb-49 30 23 restriction restriction NOUN njb-49 30 24 enzymes enzyme NOUN njb-49 30 25 were be AUX njb-49 30 26 purchased purchase VERB njb-49 30 27 from from ADP njb-49 30 28 takara takara PROPN njb-49 30 29 bio bio PROPN njb-49 30 30 . . PROPN njb-49 30 31 inc inc PROPN njb-49 30 32 . . PROPN njb-49 30 33 , , PUNCT njb-49 30 34 ( ( PUNCT njb-49 30 35 shiga shiga PROPN njb-49 30 36 , , PUNCT njb-49 30 37 japan japan PROPN njb-49 30 38 ) ) PUNCT njb-49 30 39 . . PUNCT njb-49 31 1 luria luria PROPN njb-49 31 2 bertani bertani PROPN njb-49 31 3 ( ( PUNCT njb-49 31 4 lb lb NOUN njb-49 31 5 ) ) PUNCT njb-49 31 6 media medium NOUN njb-49 31 7 and and CCONJ njb-49 31 8 cell cell NOUN njb-49 31 9 culture culture NOUN njb-49 31 10 media medium NOUN njb-49 31 11 were be AUX njb-49 31 12 purchased purchase VERB njb-49 31 13 from from ADP njb-49 31 14 becton becton PROPN njb-49 31 15 dickinson dickinson PROPN njb-49 31 16 and and CCONJ njb-49 31 17 company company PROPN njb-49 31 18 ( ( PUNCT njb-49 31 19 md md PROPN njb-49 31 20 , , PUNCT njb-49 31 21 usa usa PROPN njb-49 31 22 ) ) PUNCT njb-49 31 23 and and CCONJ njb-49 31 24 hyclone hyclone PROPN njb-49 31 25 ( ( PUNCT njb-49 31 26 ut ut PROPN njb-49 31 27 , , PUNCT njb-49 31 28 usa usa PROPN njb-49 31 29 ) ) PUNCT njb-49 31 30 respectively respectively ADV njb-49 31 31 . . PUNCT njb-49 32 1 additional additional ADJ njb-49 32 2 chemicals chemical NOUN njb-49 32 3 including include VERB njb-49 32 4 ampicillin ampicillin NOUN njb-49 32 5 , , PUNCT njb-49 32 6 iptg iptg PROPN njb-49 32 7 , , PUNCT njb-49 32 8 freund freund PROPN njb-49 32 9 ’s ’s PART njb-49 32 10 complete complete ADJ njb-49 32 11 and and CCONJ njb-49 32 12 incomplete incomplete ADJ njb-49 32 13 adjuvant adjuvant NOUN njb-49 32 14 , , PUNCT njb-49 32 15 and and CCONJ njb-49 32 16 buffers buffer NOUN njb-49 32 17 were be AUX njb-49 32 18 purchased purchase VERB njb-49 32 19 from from ADP njb-49 32 20 sigma sigma PROPN njb-49 32 21 aldrich aldrich PROPN njb-49 32 22 ( ( PUNCT njb-49 32 23 mo mo PROPN njb-49 32 24 , , PUNCT njb-49 32 25 usa usa PROPN njb-49 32 26 ) ) PUNCT njb-49 32 27 . . PUNCT njb-49 33 1 bacterial bacterial ADJ njb-49 33 2 strains strain NOUN njb-49 33 3 , , PUNCT njb-49 33 4 plasmids plasmid NOUN njb-49 33 5 , , PUNCT njb-49 33 6 and and CCONJ njb-49 33 7 purification purification NOUN njb-49 33 8 nepal nepal ADJ njb-49 33 9     SPACE njb-49 33 10 journal journal NOUN njb-49 33 11     SPACE njb-49 33 12 of of ADP njb-49 33 13     SPACE njb-49 33 14 biotechnology biotechnology NOUN njb-49 33 15 . . PUNCT njb-49 33 16         SPACE njb-49 34 1 jan jan PROPN njb-49 34 2 . . PROPN njb-49 34 3     SPACE njb-49 34 4 2012 2012 NUM njb-49 34 5 , , PUNCT njb-49 34 6     SPACE njb-49 34 7 vol vol NOUN njb-49 34 8 . . PUNCT njb-49 34 9     SPACE njb-49 34 10 2 2 NUM njb-49 34 11 , , PUNCT njb-49 34 12     SPACE njb-49 34 13 no no INTJ njb-49 34 14 . . PUNCT njb-49 34 15     SPACE njb-49 35 1 1 1 NUM njb-49 35 2 : : PUNCT njb-49 35 3     SPACE njb-49 35 4 1 1 X njb-49 35 5     SPACE njb-49 35 6 – – PUNCT njb-49 35 7     SPACE njb-49 35 8 15 15 NUM njb-49 35 9                                                                                                                                                                                                                                         SPACE njb-49 35 10 biotechnology biotechnology NOUN njb-49 35 11     SPACE njb-49 35 12 society society NOUN njb-49 35 13     SPACE njb-49 35 14 of of ADP njb-49 35 15     SPACE njb-49 35 16 nepal nepal NOUN njb-49 35 17     SPACE njb-49 35 18 ( ( PUNCT njb-49 35 19 bsn bsn PROPN njb-49 35 20 ) ) PUNCT njb-49 35 21 , , PUNCT njb-49 35 22     SPACE njb-49 35 23 all all DET njb-49 35 24     SPACE njb-49 35 25 rights right NOUN njb-49 35 26     SPACE njb-49 35 27 reserved reserve VERB njb-49 35 28     SPACE njb-49 35 29 3 3 NUM njb-49 35 30 systems system NOUN njb-49 35 31 c. c. PROPN njb-49 35 32 botulinum botulinum PROPN njb-49 35 33 type type NOUN njb-49 35 34 f f PROPN njb-49 35 35 strain strain PROPN njb-49 35 36 langeland langeland PROPN njb-49 36 1 [ [ PUNCT njb-49 36 2 23 23 NUM njb-49 36 3 ] ] PUNCT njb-49 36 4 and and CCONJ njb-49 36 5 bont bont PROPN njb-49 36 6 / / SYM njb-49 36 7 f f PROPN njb-49 36 8 ( ( PUNCT njb-49 36 9 wako wako ADJ njb-49 36 10 pure pure ADJ njb-49 36 11 chemicals chemical NOUN njb-49 36 12 ind ind NOUN njb-49 36 13 . . PROPN njb-49 36 14 , , PUNCT njb-49 36 15 osaka osaka PROPN njb-49 36 16 , , PUNCT njb-49 36 17 japan japan PROPN njb-49 36 18 ) ) PUNCT njb-49 36 19 was be AUX njb-49 36 20 kindly kindly ADV njb-49 36 21 supplied supply VERB njb-49 36 22 by by ADP njb-49 36 23 medy medy PROPN njb-49 36 24 - - PUNCT njb-49 36 25 tox tox PROPN njb-49 36 26 inc inc PROPN njb-49 36 27 . . PROPN njb-49 37 1 plasmids plasmids PROPN njb-49 37 2 pgex-4t-1 pgex-4t-1 PROPN njb-49 37 3 ( ( PUNCT njb-49 37 4 amersham amersham PROPN njb-49 37 5 pharmacia pharmacia PROPN njb-49 37 6 biotech biotech PROPN njb-49 37 7 acquired acquire VERB njb-49 37 8 by by ADP njb-49 37 9 ge ge PROPN njb-49 37 10 healthcare healthcare PROPN njb-49 37 11 , , PUNCT njb-49 37 12 uppsala uppsala PROPN njb-49 37 13 , , PUNCT njb-49 37 14 sweden sweden PROPN njb-49 37 15 ) ) PUNCT njb-49 37 16 and and CCONJ njb-49 37 17 pet-32a(+ pet-32a(+ PROPN njb-49 37 18 ) ) PUNCT njb-49 37 19 ( ( PUNCT njb-49 37 20 novagen novagen PROPN njb-49 37 21 , , PUNCT njb-49 37 22 emd emd PROPN njb-49 37 23 chemicals chemicals PROPN njb-49 37 24 inc inc PROPN njb-49 37 25 . . PROPN njb-49 37 26 , , PUNCT njb-49 37 27 affiliate affiliate NOUN njb-49 37 28 of of ADP njb-49 37 29 merck merck PROPN njb-49 37 30 kgaa kgaa PROPN njb-49 37 31 , , PUNCT njb-49 37 32 darmstadt darmstadt PROPN njb-49 37 33 , , PUNCT njb-49 37 34 germany germany PROPN njb-49 37 35 ) ) PUNCT njb-49 37 36 were be AUX njb-49 37 37 used use VERB njb-49 37 38 for for ADP njb-49 37 39 the the DET njb-49 37 40 construction construction NOUN njb-49 37 41 of of ADP njb-49 37 42 expression expression NOUN njb-49 37 43 vectors vector NOUN njb-49 37 44 . . PUNCT njb-49 38 1 e. e. PROPN njb-49 38 2 coli coli PROPN njb-49 38 3 strains strain VERB njb-49 38 4 dh5α dh5α PRON njb-49 38 5 ( ( PUNCT njb-49 38 6 takara takara PROPN njb-49 38 7 bio bio PROPN njb-49 38 8 inc inc PROPN njb-49 38 9 . . PROPN njb-49 38 10 ) ) PUNCT njb-49 38 11 , , PUNCT njb-49 38 12 bl21codonplus bl21codonplus NOUN njb-49 38 13 - - PUNCT njb-49 38 14 ril ril NOUN njb-49 38 15 and and CCONJ njb-49 38 16 bl21 bl21 NOUN njb-49 38 17 - - PUNCT njb-49 38 18 codonplus(de3)ril codonplus(de3)ril PROPN njb-49 38 19 ( ( PUNCT njb-49 38 20 stratagene stratagene NOUN njb-49 38 21 , , PUNCT njb-49 38 22 ca can AUX njb-49 38 23 , , PUNCT njb-49 38 24 usa usa PROPN njb-49 38 25 ) ) PUNCT njb-49 38 26 were be AUX njb-49 38 27 used use VERB njb-49 38 28 as as ADP njb-49 38 29 host host NOUN njb-49 38 30 strains strain NOUN njb-49 38 31 for for ADP njb-49 38 32 propagation propagation NOUN njb-49 38 33 and and CCONJ njb-49 38 34 expression expression NOUN njb-49 38 35 of of ADP njb-49 38 36 recombinant recombinant ADJ njb-49 38 37 proteins protein NOUN njb-49 38 38 , , PUNCT njb-49 38 39 respectively respectively ADV njb-49 38 40 . . PUNCT njb-49 39 1 glutathione glutathione PROPN njb-49 39 2 - - PUNCT njb-49 39 3 s s PROPN njb-49 39 4 - - PUNCT njb-49 39 5 transferase transferase NOUN njb-49 39 6 ( ( PUNCT njb-49 39 7 gst gst NOUN njb-49 39 8 ) ) PUNCT njb-49 39 9 purification purification NOUN njb-49 39 10 modules module NOUN njb-49 39 11 and and CCONJ njb-49 39 12 thrombin thrombin NOUN njb-49 39 13 protease protease NOUN njb-49 39 14 were be AUX njb-49 39 15 purchased purchase VERB njb-49 39 16 from from ADP njb-49 39 17 amersham amersham PROPN njb-49 39 18 pharmacia pharmacia PROPN njb-49 39 19 biotech biotech PROPN njb-49 39 20 . . PUNCT njb-49 40 1 construction construction NOUN njb-49 40 2 of of ADP njb-49 40 3 bont bont PROPN njb-49 40 4 / / SYM njb-49 40 5 f f PROPN njb-49 40 6 domains domain NOUN njb-49 40 7 chromosomal chromosomal ADJ njb-49 40 8 dna dna PROPN njb-49 40 9 was be AUX njb-49 40 10 isolated isolate VERB njb-49 40 11 from from ADP njb-49 40 12 c. c. PROPN njb-49 40 13 botulinum botulinum PROPN njb-49 40 14 f f PROPN njb-49 40 15 str str PROPN njb-49 40 16 . . PUNCT njb-49 41 1 langeland langeland PROPN njb-49 41 2 , , PUNCT njb-49 41 3 which which PRON njb-49 41 4 was be AUX njb-49 41 5 used use VERB njb-49 41 6 as as ADP njb-49 41 7 the the DET njb-49 41 8 template template NOUN njb-49 41 9 for for ADP njb-49 41 10 polymerase polymerase NOUN njb-49 41 11 chain chain NOUN njb-49 41 12 reaction reaction NOUN njb-49 41 13 ( ( PUNCT njb-49 41 14 pcr pcr PROPN njb-49 41 15 ) ) PUNCT njb-49 41 16 for for ADP njb-49 41 17 all all DET njb-49 41 18 domains domain NOUN njb-49 41 19 ( ( PUNCT njb-49 41 20 fig fig NOUN njb-49 41 21 . . PROPN njb-49 41 22 1 1 NUM njb-49 41 23 ) ) PUNCT njb-49 41 24 . . PUNCT njb-49 42 1 forward forward ADV njb-49 42 2 and and CCONJ njb-49 42 3 reverse reverse VERB njb-49 42 4 primers primer NOUN njb-49 42 5 for for SCONJ njb-49 42 6 each each DET njb-49 42 7 domain domain NOUN njb-49 42 8 were be AUX njb-49 42 9 designed design VERB njb-49 42 10 ( ( PUNCT njb-49 42 11 table table NOUN njb-49 42 12 1 1 NUM njb-49 42 13 ) ) PUNCT njb-49 42 14 based base VERB njb-49 42 15 on on ADP njb-49 42 16 the the DET njb-49 42 17 published publish VERB njb-49 42 18 ncbi ncbi NOUN njb-49 42 19 sequence sequence NOUN njb-49 42 20 of of ADP njb-49 42 21 the the DET njb-49 42 22 langeland langeland PROPN njb-49 42 23 genome genome NOUN njb-49 42 24 ( ( PUNCT njb-49 42 25 nc_009699 nc_009699 PROPN njb-49 42 26 ) ) PUNCT njb-49 42 27 . . PUNCT njb-49 43 1 synthetic synthetic ADJ njb-49 43 2 genes gene NOUN njb-49 43 3 encoding encode VERB njb-49 43 4 bont bont PROPN njb-49 43 5 / / SYM njb-49 43 6 f f PROPN njb-49 43 7 lc lc PROPN njb-49 43 8 ( ( PUNCT njb-49 43 9 light light ADJ njb-49 43 10 chain chain NOUN njb-49 43 11 ) ) PUNCT njb-49 43 12 , , PUNCT njb-49 43 13 catalytic catalytic ADJ njb-49 43 14 domain domain NOUN njb-49 43 15 ( ( PUNCT njb-49 43 16 1308 1308 NUM njb-49 43 17 bp bp PROPN njb-49 43 18 ) ) PUNCT njb-49 43 19 ; ; PUNCT njb-49 43 20 hn hn PROPN njb-49 43 21 ( ( PUNCT njb-49 43 22 n n ADJ njb-49 43 23 - - ADJ njb-49 43 24 terminal terminal ADJ njb-49 43 25 1/2 1/2 NUM njb-49 43 26 of of ADP njb-49 43 27 heavy heavy ADJ njb-49 43 28 chain chain NOUN njb-49 43 29 ) ) PUNCT njb-49 43 30 , , PUNCT njb-49 43 31 translocation translocation NOUN njb-49 43 32 domain domain NOUN njb-49 43 33 ( ( PUNCT njb-49 43 34 1266 1266 NUM njb-49 43 35 bp bp PROPN njb-49 43 36 ) ) PUNCT njb-49 43 37 ; ; PUNCT njb-49 44 1 hc hc INTJ njb-49 44 2 ( ( PUNCT njb-49 44 3 c c NOUN njb-49 44 4 - - PUNCT njb-49 44 5 terminal terminal NOUN njb-49 44 6 1/2 1/2 NUM njb-49 44 7 of of ADP njb-49 44 8 heavy heavy ADJ njb-49 44 9 chain chain NOUN njb-49 44 10 ) ) PUNCT njb-49 44 11 , , PUNCT njb-49 44 12 receptor receptor NOUN njb-49 44 13 binding bind VERB njb-49 44 14 domain domain NOUN njb-49 44 15 ( ( PUNCT njb-49 44 16 1263 1263 NUM njb-49 44 17 bp bp PROPN njb-49 44 18 ) ) PUNCT njb-49 44 19 ; ; PUNCT njb-49 44 20 and and CCONJ njb-49 44 21 hcc hcc PROPN njb-49 44 22 ( ( PUNCT njb-49 44 23 far far ADV njb-49 44 24 cterminal cterminal ADJ njb-49 44 25 quarter quarter NOUN njb-49 44 26 part part NOUN njb-49 44 27 of of ADP njb-49 44 28 heavy heavy ADJ njb-49 44 29 chain chain NOUN njb-49 44 30 ) ) PUNCT njb-49 44 31 , , PUNCT njb-49 44 32 ( ( PUNCT njb-49 44 33 501 501 NUM njb-49 44 34 bp bp PROPN njb-49 44 35 ) ) PUNCT njb-49 44 36 were be AUX njb-49 44 37 designed design VERB njb-49 44 38 based base VERB njb-49 44 39 on on ADP njb-49 44 40 the the DET njb-49 44 41 published publish VERB njb-49 44 42 bont bont ADJ njb-49 44 43 / / SYM njb-49 44 44 f f PROPN njb-49 44 45 sequence sequence NOUN njb-49 44 46 ( ( PUNCT njb-49 44 47 orf orf NOUN njb-49 44 48 : : PUNCT njb-49 44 49 883635 883635 NUM njb-49 44 50 - - SYM njb-49 44 51 887471 887471 NUM njb-49 44 52 ) ) PUNCT njb-49 44 53 langeland langeland PROPN njb-49 44 54 str str PROPN njb-49 44 55 . . PUNCT njb-49 45 1 ( ( PUNCT njb-49 45 2 ncbi ncbi PROPN njb-49 45 3 genbank genbank PROPN njb-49 45 4 accession accession PROPN njb-49 45 5 : : PUNCT njb-49 45 6 nc_009699 nc_009699 PROPN njb-49 45 7 ) ) PUNCT njb-49 45 8 for for ADP njb-49 45 9 the the DET njb-49 45 10 plasmid plasmid ADJ njb-49 45 11 construction construction NOUN njb-49 45 12 of of ADP njb-49 45 13 each each DET njb-49 45 14 domain domain NOUN njb-49 45 15 . . PUNCT njb-49 46 1 bp bp PROPN njb-49 46 2 indicates indicate VERB njb-49 46 3 base base NOUN njb-49 46 4 pair pair NOUN njb-49 46 5 of of ADP njb-49 46 6 dna dna PROPN njb-49 46 7 . . PUNCT njb-49 47 1 1 1 NUM njb-49 47 2 kb kb PROPN njb-49 47 3 marker marker NOUN njb-49 47 4 is be AUX njb-49 47 5 provided provide VERB njb-49 47 6 for for ADP njb-49 47 7 scale scale NOUN njb-49 47 8 . . PUNCT njb-49 48 1 table table NOUN njb-49 48 2 1 1 NUM njb-49 48 3 . . PUNCT njb-49 49 1 primers primer NOUN njb-49 49 2 used use VERB njb-49 49 3 for for ADP njb-49 49 4 amplification amplification NOUN njb-49 49 5 of of ADP njb-49 49 6 bont bont PROPN njb-49 49 7 / / SYM njb-49 49 8 f f PROPN njb-49 49 9 domains domain NOUN njb-49 49 10 . . PUNCT njb-49 50 1 aprimer aprimer PROPN njb-49 50 2 direction direction NOUN njb-49 50 3 : : PUNCT njb-49 50 4 f f X njb-49 50 5 , , PUNCT njb-49 50 6 forward forward ADV njb-49 50 7 ; ; PUNCT njb-49 50 8 r r X njb-49 50 9 , , PUNCT njb-49 50 10 reverse reverse NOUN njb-49 50 11 . . PUNCT njb-49 51 1 bdirection bdirection NOUN njb-49 51 2 of of ADP njb-49 51 3 each each DET njb-49 51 4 sequence sequence NOUN njb-49 51 5 is be AUX njb-49 51 6 in in ADP njb-49 51 7 5 5 NUM njb-49 51 8 ' ' PUNCT njb-49 51 9 to to ADP njb-49 51 10 3 3 PROPN njb-49 51 11 ' ' PUNCT njb-49 51 12 orientation orientation NOUN njb-49 51 13 ( ( PUNCT njb-49 51 14 underlined underline VERB njb-49 51 15 ) ) PUNCT njb-49 51 16 including include VERB njb-49 51 17 restriction restriction NOUN njb-49 51 18 endonuclease endonuclease NOUN njb-49 51 19 sequence sequence NOUN njb-49 51 20 ( ( PUNCT njb-49 51 21 bold bold ADJ njb-49 51 22 ) ) PUNCT njb-49 51 23 and and CCONJ njb-49 51 24 inserted insert VERB njb-49 51 25 stop stop NOUN njb-49 51 26 codon codon NOUN njb-49 51 27 sequences sequence NOUN njb-49 51 28 ( ( PUNCT njb-49 51 29 blue blue NOUN njb-49 51 30 ) ) PUNCT njb-49 51 31 for for ADP njb-49 51 32 lc lc NOUN njb-49 51 33 and and CCONJ njb-49 51 34 hn hn NOUN njb-49 51 35 reverse reverse ADJ njb-49 51 36 primers primer NOUN njb-49 51 37 . . PUNCT njb-49 52 1 cnucleotide cnucleotide PROPN njb-49 52 2 positions position NOUN njb-49 52 3 based base VERB njb-49 52 4 on on ADP njb-49 52 5 published publish VERB njb-49 52 6 c. c. PROPN njb-49 52 7 botulinum botulinum PROPN njb-49 52 8 f f PROPN njb-49 52 9 str str PROPN njb-49 52 10 . . PUNCT njb-49 53 1 langeland langeland PROPN njb-49 53 2 genome genome NOUN njb-49 53 3 sequence sequence NOUN njb-49 53 4 ( ( PUNCT njb-49 53 5 ncbi ncbi PROPN njb-49 53 6 genbank genbank PROPN njb-49 53 7 accession accession PROPN njb-49 53 8 : : PUNCT njb-49 53 9 nc_009699 nc_009699 PROPN njb-49 53 10 ) ) PUNCT njb-49 53 11 . . PUNCT njb-49 54 1 drestriction drestriction NOUN njb-49 54 2 endonucleases endonuclease NOUN njb-49 54 3 utilized utilize VERB njb-49 54 4 to to PART njb-49 54 5 digest digest VERB njb-49 54 6 pcr pcr NOUN njb-49 54 7 amplified amplify VERB njb-49 54 8 domain domain NOUN njb-49 54 9 and and CCONJ njb-49 54 10 vector vector NOUN njb-49 54 11 for for ADP njb-49 54 12 plasmid plasmid ADJ njb-49 54 13 construction construction NOUN njb-49 54 14 . . PUNCT njb-49 55 1 isolation isolation NOUN njb-49 55 2 of of ADP njb-49 55 3 genomic genomic ADJ njb-49 55 4 dna dna PROPN njb-49 55 5 was be AUX njb-49 55 6 done do VERB njb-49 55 7 utilizing utilize VERB njb-49 55 8 protocols protocol NOUN njb-49 55 9 and and CCONJ njb-49 55 10 chemicals chemical NOUN njb-49 55 11 mentioned mention VERB njb-49 55 12 by by ADP njb-49 55 13 wizard wizard NOUN njb-49 55 14 genomic genomic ADJ njb-49 55 15 dna dna PROPN njb-49 55 16 purification purification NOUN njb-49 55 17 kit kit NOUN njb-49 55 18 ( ( PUNCT njb-49 55 19 promega promega PROPN njb-49 55 20 , , PUNCT njb-49 55 21 wi wi PROPN njb-49 55 22 , , PUNCT njb-49 55 23 usa usa PROPN njb-49 55 24 ) ) PUNCT njb-49 55 25 . . PUNCT njb-49 56 1 pcr pcr PROPN njb-49 56 2 was be AUX njb-49 56 3 performed perform VERB njb-49 56 4 using use VERB njb-49 56 5 a a DET njb-49 56 6 bio bio ADJ njb-49 56 7 - - ADJ njb-49 56 8 rad rad ADJ njb-49 56 9 icycler icycler NOUN njb-49 56 10 ( ( PUNCT njb-49 56 11 ca ca PROPN njb-49 56 12 , , PUNCT njb-49 56 13 usa usa PROPN njb-49 56 14 ) ) PUNCT njb-49 56 15 . . PUNCT njb-49 57 1 reaction reaction NOUN njb-49 57 2 mixtures mixture NOUN njb-49 57 3 were be AUX njb-49 57 4 preheated preheat VERB njb-49 57 5 for for ADP njb-49 57 6 5 5 NUM njb-49 57 7 min min NOUN njb-49 57 8 at at ADP njb-49 57 9 94 94 NUM njb-49 57 10 ° ° PUNCT njb-49 57 11 c c NOUN njb-49 57 12 for for ADP njb-49 57 13 initial initial ADJ njb-49 57 14 denaturation denaturation NOUN njb-49 57 15 and and CCONJ njb-49 57 16 then then ADV njb-49 57 17 30 30 NUM njb-49 57 18 cycles cycle NOUN njb-49 57 19 of of ADP njb-49 57 20 pcr pcr NOUN njb-49 57 21 ( ( PUNCT njb-49 57 22 denaturation denaturation NOUN njb-49 57 23 , , PUNCT njb-49 57 24 annealing annealing NOUN njb-49 57 25 , , PUNCT njb-49 57 26 and and CCONJ njb-49 57 27 elongation elongation NOUN njb-49 57 28 ) ) PUNCT njb-49 57 29 were be AUX njb-49 57 30 performed perform VERB njb-49 57 31 for for ADP njb-49 57 32 amplification amplification NOUN njb-49 57 33 of of ADP njb-49 57 34 domains domain NOUN njb-49 57 35 as as SCONJ njb-49 57 36 follows follow VERB njb-49 57 37 : : PUNCT njb-49 57 38 1 1 NUM njb-49 57 39 min min NOUN njb-49 57 40 at at ADP njb-49 57 41 94 94 NUM njb-49 57 42 ° ° PUNCT njb-49 57 43 c c NOUN njb-49 57 44 , , PUNCT njb-49 57 45 1 1 NUM njb-49 57 46 min min NOUN njb-49 57 47 at at ADP njb-49 57 48 57 57 NUM njb-49 57 49 ° ° NOUN njb-49 57 50 c c NOUN njb-49 57 51 , , PUNCT njb-49 57 52 and and CCONJ njb-49 57 53 1 1 NUM njb-49 57 54 min min NOUN njb-49 57 55 30 30 NUM njb-49 57 56 sec sec NOUN njb-49 57 57 at at ADP njb-49 57 58 72 72 NUM njb-49 57 59 ° ° NOUN njb-49 57 60 c c NOUN njb-49 57 61 for for ADP njb-49 57 62 f f PROPN njb-49 57 63 - - PUNCT njb-49 57 64 lc lc PROPN njb-49 57 65 ; ; PUNCT njb-49 57 66 1 1 NUM njb-49 57 67 min min NOUN njb-49 57 68 at at ADP njb-49 57 69 94 94 NUM njb-49 57 70 ° ° PUNCT njb-49 57 71 c c NOUN njb-49 57 72 , , PUNCT njb-49 57 73 1 1 NUM njb-49 57 74 min min NOUN njb-49 57 75 at at ADP njb-49 57 76 50 50 NUM njb-49 57 77 ° ° NOUN njb-49 57 78 c c NOUN njb-49 57 79 , , PUNCT njb-49 57 80 and and CCONJ njb-49 57 81 1 1 NUM njb-49 57 82 min min NOUN njb-49 57 83 30 30 NUM njb-49 57 84 sec sec NOUN njb-49 57 85 at at ADP njb-49 57 86 72 72 NUM njb-49 57 87 ° ° NOUN njb-49 57 88 c c NOUN njb-49 57 89 for for ADP njb-49 57 90 f f PROPN njb-49 57 91 - - PROPN njb-49 57 92 hn hn PROPN njb-49 57 93 and and CCONJ njb-49 57 94 f f X njb-49 57 95 - - PUNCT njb-49 57 96 hc hc PROPN njb-49 57 97 ; ; PUNCT njb-49 57 98 1 1 NUM njb-49 57 99 min min NOUN njb-49 57 100 at at ADP njb-49 57 101 94 94 NUM njb-49 57 102 ° ° PUNCT njb-49 57 103 c c NOUN njb-49 57 104 , , PUNCT njb-49 57 105 1 1 NUM njb-49 57 106 min min NOUN njb-49 57 107 at at ADP njb-49 57 108 54.5 54.5 NUM njb-49 57 109 ° ° NOUN njb-49 57 110 c c NOUN njb-49 57 111 , , PUNCT njb-49 57 112 and and CCONJ njb-49 57 113 40 40 NUM njb-49 57 114 sec sec NOUN njb-49 57 115 at at ADP njb-49 57 116 72 72 NUM njb-49 57 117 ° ° NOUN njb-49 57 118 c c NOUN njb-49 57 119 for for ADP njb-49 57 120 f f PROPN njb-49 57 121 - - PUNCT njb-49 57 122 hcc hcc PROPN njb-49 57 123 . . PUNCT njb-49 58 1 following follow VERB njb-49 58 2 completion completion NOUN njb-49 58 3 of of ADP njb-49 58 4 cycles cycle NOUN njb-49 58 5 , , PUNCT njb-49 58 6 a a DET njb-49 58 7 final final ADJ njb-49 58 8 extension extension NOUN njb-49 58 9 was be AUX njb-49 58 10 lc lc NOUN njb-49 58 11 1 1 NUM njb-49 58 12 kb kb PROPN njb-49 58 13 scale scale PROPN njb-49 58 14 s s PROPN njb-49 58 15 s s PROPN njb-49 59 1 hn hn PROPN njb-49 60 1 hc hc NOUN njb-49 60 2 1308 1308 NUM njb-49 61 1 bp bp PROPN njb-49 61 2 1266 1266 NUM njb-49 61 3 bp bp PROPN njb-49 61 4 1263 1263 NUM njb-49 61 5 bp bp PROPN njb-49 61 6 hcc hcc PROPN njb-49 61 7 501 501 NUM njb-49 61 8 bp bp PROPN njb-49 61 9 native native PROPN njb-49 61 10 bont bont PROPN njb-49 61 11 / / SYM njb-49 61 12 f f PROPN njb-49 61 13 ( ( PUNCT njb-49 61 14 3837 3837 NUM njb-49 61 15 bp bp PROPN njb-49 61 16 ) ) PUNCT njb-49 61 17 883635 883635 NUM njb-49 61 18 887471 887471 NUM njb-49 61 19 _ _ PUNCT njb-49 61 20 _ _ PUNCT njb-49 61 21 _ _ PUNCT njb-49 61 22 _ _ PUNCT njb-49 61 23 _ _ PUNCT njb-49 61 24 _ _ PUNCT njb-49 61 25 _ _ PUNCT njb-49 61 26 _ _ PUNCT njb-49 61 27 _ _ PUNCT njb-49 61 28 _ _ PUNCT njb-49 61 29 _ _ PUNCT njb-49 61 30 _ _ PUNCT njb-49 61 31 _ _ PUNCT njb-49 61 32 _ _ PUNCT njb-49 61 33 _ _ PUNCT njb-49 61 34 _ _ PUNCT njb-49 61 35 _ _ PUNCT njb-49 61 36 _ _ PUNCT njb-49 61 37 _ _ PUNCT njb-49 61 38 _ _ PUNCT njb-49 61 39 _ _ PUNCT njb-49 61 40 _ _ PUNCT njb-49 61 41 _ _ PUNCT njb-49 61 42 _ _ PUNCT njb-49 61 43 _ _ PUNCT njb-49 61 44 _ _ PUNCT njb-49 61 45 _ _ PUNCT njb-49 61 46 _ _ PUNCT njb-49 61 47 _ _ PUNCT njb-49 61 48 _ _ PUNCT njb-49 61 49 _ _ PUNCT njb-49 61 50 _ _ PUNCT njb-49 61 51 _ _ PUNCT njb-49 61 52 _ _ PUNCT njb-49 61 53 _ _ PUNCT njb-49 61 54 _ _ PUNCT njb-49 61 55 _ _ PUNCT njb-49 61 56 _ _ PUNCT njb-49 61 57 _ _ PUNCT njb-49 61 58 _ _ PUNCT njb-49 61 59 _ _ PUNCT njb-49 61 60 _ _ PUNCT njb-49 61 61 _ _ PUNCT njb-49 61 62 _ _ PUNCT njb-49 61 63 _ _ PUNCT njb-49 61 64 _ _ PUNCT njb-49 61 65 _ _ PUNCT njb-49 61 66 _ _ PUNCT njb-49 61 67 _ _ PUNCT njb-49 61 68 _ _ PUNCT njb-49 61 69 _ _ PUNCT njb-49 61 70 _ _ PUNCT njb-49 61 71 _ _ PUNCT njb-49 61 72 _ _ PUNCT njb-49 61 73 _ _ PUNCT njb-49 61 74 _ _ PUNCT njb-49 61 75 _ _ PUNCT njb-49 61 76 _ _ PUNCT njb-49 61 77 _ _ PUNCT njb-49 61 78 _ _ PUNCT njb-49 61 79 _ _ PUNCT njb-49 61 80 _ _ PUNCT njb-49 61 81 _ _ PUNCT njb-49 61 82 _ _ PUNCT njb-49 61 83 _ _ PUNCT njb-49 61 84 _ _ PUNCT njb-49 61 85 _ _ PUNCT njb-49 61 86 _ _ PUNCT njb-49 61 87 _ _ PUNCT njb-49 61 88 _ _ PUNCT njb-49 61 89 _ _ PUNCT njb-49 61 90 _ _ PUNCT njb-49 61 91 _ _ PUNCT njb-49 61 92 _ _ PUNCT njb-49 61 93 _ _ PUNCT njb-49 61 94 _ _ PUNCT njb-49 61 95 _ _ PUNCT njb-49 61 96 _ _ PUNCT njb-49 61 97 _ _ PUNCT njb-49 61 98 _ _ PUNCT njb-49 61 99 _ _ PUNCT njb-49 61 100 _ _ PUNCT njb-49 61 101 _ _ PUNCT njb-49 61 102 _ _ PUNCT njb-49 61 103 _ _ PUNCT njb-49 61 104 _ _ PUNCT njb-49 61 105 _ _ PUNCT njb-49 61 106 _ _ PUNCT njb-49 61 107 _ _ PUNCT njb-49 61 108 _ _ PUNCT njb-49 61 109 _ _ PUNCT njb-49 61 110 _ _ PUNCT njb-49 61 111 _ _ PUNCT njb-49 61 112 _ _ PUNCT njb-49 61 113 _ _ PUNCT njb-49 61 114 _ _ PUNCT njb-49 61 115 _ _ PUNCT njb-49 61 116 _ _ PUNCT njb-49 61 117 _ _ PUNCT njb-49 61 118 _ _ PUNCT njb-49 61 119 _ _ PUNCT njb-49 61 120 _ _ PUNCT njb-49 61 121 _ _ PUNCT njb-49 61 122 _ _ PUNCT njb-49 61 123 _ _ PUNCT njb-49 61 124 _ _ PUNCT njb-49 61 125 _ _ PUNCT njb-49 61 126 _ _ PUNCT njb-49 61 127 _ _ PUNCT njb-49 61 128 _ _ PUNCT njb-49 61 129 _ _ PUNCT njb-49 61 130 _ _ PUNCT njb-49 61 131 _ _ PUNCT njb-49 61 132 _ _ PUNCT njb-49 61 133 _ _ PUNCT njb-49 61 134 _ _ PUNCT njb-49 61 135 _ _ PUNCT njb-49 61 136 _ _ PUNCT njb-49 61 137 _ _ PUNCT njb-49 61 138 _ _ PUNCT njb-49 61 139 _ _ PUNCT njb-49 61 140 _ _ PUNCT njb-49 61 141 _ _ PUNCT njb-49 61 142 _ _ PUNCT njb-49 61 143 _ _ PUNCT njb-49 62 1 primera primera PROPN njb-49 62 2 sequence sequence NOUN njb-49 62 3 5 5 NUM njb-49 62 4 ' ' PUNCT njb-49 62 5 ! ! PUNCT njb-49 63 1 3'b 3'b NOUN njb-49 63 2 nucleotide nucleotide NOUN njb-49 63 3 positionsc positionsc PROPN njb-49 63 4 restriction restriction NOUN njb-49 63 5 enzymesd enzymesd VERB njb-49 63 6 _ _ PUNCT njb-49 63 7 _ _ PUNCT njb-49 63 8 _ _ PUNCT njb-49 63 9 _ _ PUNCT njb-49 63 10 _ _ PUNCT njb-49 63 11 _ _ PUNCT njb-49 63 12 _ _ PUNCT njb-49 63 13 _ _ PUNCT njb-49 63 14 _ _ PUNCT njb-49 63 15 _ _ PUNCT njb-49 63 16 _ _ PUNCT njb-49 63 17 _ _ PUNCT njb-49 63 18 _ _ PUNCT njb-49 63 19 _ _ PUNCT njb-49 63 20 _ _ PUNCT njb-49 63 21 _ _ PUNCT njb-49 63 22 _ _ PUNCT njb-49 63 23 _ _ PUNCT njb-49 63 24 _ _ PUNCT njb-49 63 25 _ _ PUNCT njb-49 63 26 _ _ PUNCT njb-49 63 27 _ _ PUNCT njb-49 63 28 _ _ PUNCT njb-49 63 29 _ _ PUNCT njb-49 63 30 _ _ PUNCT njb-49 63 31 _ _ PUNCT njb-49 63 32 _ _ PUNCT njb-49 63 33 _ _ PUNCT njb-49 63 34 _ _ PUNCT njb-49 63 35 _ _ PUNCT njb-49 63 36 _ _ PUNCT njb-49 63 37 _ _ PUNCT njb-49 63 38 _ _ PUNCT njb-49 63 39 _ _ PUNCT njb-49 63 40 _ _ PUNCT njb-49 63 41 _ _ PUNCT njb-49 63 42 _ _ PUNCT njb-49 63 43 _ _ PUNCT njb-49 63 44 _ _ PUNCT njb-49 63 45 _ _ PUNCT njb-49 63 46 _ _ PUNCT njb-49 63 47 _ _ PUNCT njb-49 63 48 _ _ PUNCT njb-49 63 49 _ _ PUNCT njb-49 63 50 _ _ PUNCT njb-49 63 51 _ _ PUNCT njb-49 63 52 _ _ PUNCT njb-49 63 53 _ _ PUNCT njb-49 63 54 _ _ PUNCT njb-49 63 55 _ _ PUNCT njb-49 63 56 _ _ PUNCT njb-49 63 57 _ _ PUNCT njb-49 63 58 _ _ PUNCT njb-49 63 59 _ _ PUNCT njb-49 63 60 _ _ PUNCT njb-49 63 61 _ _ PUNCT njb-49 63 62 _ _ PUNCT njb-49 63 63 _ _ PUNCT njb-49 63 64 _ _ PUNCT njb-49 63 65 _ _ PUNCT njb-49 63 66 _ _ PUNCT njb-49 63 67 _ _ PUNCT njb-49 63 68 _ _ PUNCT njb-49 63 69 _ _ PUNCT njb-49 63 70 _ _ PUNCT njb-49 63 71 _ _ PUNCT njb-49 63 72 _ _ PUNCT njb-49 63 73 _ _ PUNCT njb-49 63 74 _ _ PUNCT njb-49 63 75 _ _ PUNCT njb-49 63 76 _ _ PUNCT njb-49 63 77 _ _ PUNCT njb-49 63 78 _ _ PUNCT njb-49 63 79 _ _ PUNCT njb-49 63 80 _ _ PUNCT njb-49 63 81 _ _ PUNCT njb-49 63 82 _ _ PUNCT njb-49 63 83 _ _ PUNCT njb-49 63 84 _ _ PUNCT njb-49 63 85 _ _ PUNCT njb-49 63 86 _ _ PUNCT njb-49 63 87 _ _ PUNCT njb-49 63 88 _ _ PUNCT njb-49 63 89 _ _ PUNCT njb-49 63 90 _ _ PUNCT njb-49 63 91 _ _ PUNCT njb-49 63 92 _ _ PUNCT njb-49 63 93 _ _ PUNCT njb-49 63 94 _ _ PUNCT njb-49 63 95 _ _ PUNCT njb-49 63 96 _ _ PUNCT njb-49 63 97 _ _ PUNCT njb-49 63 98 _ _ PUNCT njb-49 63 99 _ _ PUNCT njb-49 63 100 _ _ PUNCT njb-49 63 101 _ _ PUNCT njb-49 63 102 _ _ PUNCT njb-49 63 103 _ _ PUNCT njb-49 63 104 _ _ PUNCT njb-49 63 105 _ _ PUNCT njb-49 63 106 _ _ PUNCT njb-49 63 107 _ _ PUNCT njb-49 63 108 _ _ PUNCT njb-49 63 109 _ _ PUNCT njb-49 63 110 _ _ PUNCT njb-49 63 111 _ _ PUNCT njb-49 63 112 _ _ PUNCT njb-49 63 113 _ _ PUNCT njb-49 63 114 _ _ PUNCT njb-49 63 115 _ _ PUNCT njb-49 63 116 _ _ PUNCT njb-49 63 117 _ _ PUNCT njb-49 63 118 _ _ PUNCT njb-49 63 119 _ _ PUNCT njb-49 63 120 _ _ PUNCT njb-49 63 121 _ _ PUNCT njb-49 63 122 _ _ PUNCT njb-49 63 123 _ _ PUNCT njb-49 63 124 _ _ PUNCT njb-49 63 125 _ _ PUNCT njb-49 63 126 _ _ PUNCT njb-49 63 127 _ _ PUNCT njb-49 63 128 _ _ PUNCT njb-49 63 129 _ _ PUNCT njb-49 63 130 _ _ PUNCT njb-49 64 1 f f X njb-49 64 2 - - PUNCT njb-49 64 3 lc lc PROPN njb-49 64 4 - - PUNCT njb-49 64 5 f f PROPN njb-49 64 6 cgtgtcgacatgccagttgtaataaatagtttt cgtgtcgacatgccagttgtaataaatagtttt PROPN njb-49 64 7 883635 883635 NUM njb-49 64 8 - - SYM njb-49 64 9 883658 883658 NUM njb-49 64 10 sali sali ADJ njb-49 64 11 f f X njb-49 64 12 - - PUNCT njb-49 64 13 lc lc NOUN njb-49 64 14 - - PUNCT njb-49 64 15 r r NOUN njb-49 64 16 ccggcggccgcttattttctaggaataacgctctt ccggcggccgcttattttctaggaataacgctctt NOUN njb-49 64 17 884942 884942 NUM njb-49 64 18 - - SYM njb-49 64 19 884922 884922 NUM njb-49 64 20 noti noti PROPN njb-49 65 1 f f X njb-49 65 2 - - PUNCT njb-49 65 3 hn hn PROPN njb-49 65 4 - - PUNCT njb-49 65 5 f f PROPN njb-49 65 6 cgtggatccggtacaaaggcgccaccgcgacta cgtggatccggtacaaaggcgccaccgcgacta PROPN njb-49 65 7 884943 884943 NUM njb-49 65 8 - - SYM njb-49 65 9 884966 884966 NUM njb-49 65 10 bamhi bamhi VERB njb-49 65 11 f f X njb-49 65 12 - - PUNCT njb-49 65 13 hn hn PROPN njb-49 65 14 - - PUNCT njb-49 65 15 r r PROPN njb-49 65 16 ccgctcgagttaatataaaattagaattttatc ccgctcgagttaatataaaattagaattttatc PROPN njb-49 65 17 886209 886209 NUM njb-49 65 18 - - PUNCT njb-49 65 19 886188 886188 NUM njb-49 65 20 xhoi xhoi PROPN njb-49 65 21 f f X njb-49 65 22 - - PUNCT njb-49 65 23 hc hc PROPN njb-49 65 24 - - PUNCT njb-49 65 25 f f PROPN njb-49 65 26 cgtggatcctttaataaattatataaaaaaatt cgtggatcctttaataaattatataaaaaaatt PROPN njb-49 65 27 886209 886209 NUM njb-49 65 28 - - PUNCT njb-49 65 29 886232 886232 NUM njb-49 65 30 bamhi bamhi VERB njb-49 65 31 f f X njb-49 65 32 - - PUNCT njb-49 65 33 hc hc NOUN njb-49 65 34 - - PUNCT njb-49 65 35 r r NOUN njb-49 65 36 ccgctcgagttagttttcttgccatccatgctc ccgctcgagttagttttcttgccatccatgctc NOUN njb-49 65 37 887471 887471 NUM njb-49 65 38 - - SYM njb-49 65 39 887448 887448 NUM njb-49 65 40 xhoi xhoi PROPN njb-49 65 41 f f PROPN njb-49 65 42 - - PUNCT njb-49 65 43 hcc hcc PROPN njb-49 65 44 - - PUNCT njb-49 65 45 f f PROPN njb-49 65 46 cgtggatccgataagtctattactcagaattca cgtggatccgataagtctattactcagaattca PROPN njb-49 65 47 886971 886971 NUM njb-49 65 48 - - SYM njb-49 65 49 886994 886994 NUM njb-49 65 50 bamhi bamhi VERB njb-49 65 51 f f X njb-49 65 52 - - PUNCT njb-49 65 53 hcc hcc PROPN njb-49 65 54 - - PUNCT njb-49 65 55 r r NOUN njb-49 65 56 ccgctcgagttagttttcttgccatccatgctc ccgctcgagttagttttcttgccatccatgctc NOUN njb-49 65 57 887471 887471 NUM njb-49 65 58 - - SYM njb-49 65 59 887448 887448 NUM njb-49 65 60 xhoi xhoi PROPN njb-49 65 61 _ _ PUNCT njb-49 65 62 _ _ PUNCT njb-49 65 63 _ _ PUNCT njb-49 65 64 _ _ PUNCT njb-49 65 65 _ _ PUNCT njb-49 65 66 _ _ PUNCT njb-49 65 67 _ _ PUNCT njb-49 65 68 _ _ PUNCT njb-49 65 69 _ _ PUNCT njb-49 65 70 _ _ PUNCT njb-49 65 71 _ _ PUNCT njb-49 65 72 _ _ PUNCT njb-49 65 73 _ _ PUNCT njb-49 65 74 _ _ PUNCT njb-49 65 75 _ _ PUNCT njb-49 65 76 _ _ PUNCT njb-49 65 77 _ _ PUNCT njb-49 65 78 _ _ PUNCT njb-49 65 79 _ _ PUNCT njb-49 65 80 _ _ PUNCT njb-49 65 81 _ _ PUNCT njb-49 65 82 _ _ PUNCT njb-49 65 83 _ _ PUNCT njb-49 65 84 _ _ PUNCT njb-49 65 85 _ _ PUNCT njb-49 65 86 _ _ PUNCT njb-49 65 87 _ _ PUNCT njb-49 65 88 _ _ PUNCT njb-49 65 89 _ _ PUNCT njb-49 65 90 _ _ PUNCT njb-49 65 91 _ _ PUNCT njb-49 65 92 _ _ PUNCT njb-49 65 93 _ _ PUNCT njb-49 65 94 _ _ PUNCT njb-49 65 95 _ _ PUNCT njb-49 65 96 _ _ PUNCT njb-49 65 97 _ _ PUNCT njb-49 65 98 _ _ PUNCT njb-49 65 99 _ _ PUNCT njb-49 65 100 _ _ PUNCT njb-49 65 101 _ _ PUNCT njb-49 65 102 _ _ PUNCT njb-49 65 103 _ _ PUNCT njb-49 65 104 _ _ PUNCT njb-49 65 105 _ _ PUNCT njb-49 65 106 _ _ PUNCT njb-49 65 107 _ _ PUNCT njb-49 65 108 _ _ PUNCT njb-49 65 109 _ _ PUNCT njb-49 65 110 _ _ PUNCT njb-49 65 111 _ _ PUNCT njb-49 65 112 _ _ PUNCT njb-49 65 113 _ _ PUNCT njb-49 65 114 _ _ PUNCT njb-49 65 115 _ _ PUNCT njb-49 65 116 _ _ PUNCT njb-49 65 117 _ _ PUNCT njb-49 65 118 _ _ PUNCT njb-49 65 119 _ _ PUNCT njb-49 65 120 _ _ PUNCT njb-49 65 121 _ _ PUNCT njb-49 65 122 _ _ PUNCT njb-49 65 123 _ _ PUNCT njb-49 65 124 _ _ PUNCT njb-49 65 125 _ _ PUNCT njb-49 65 126 _ _ PUNCT njb-49 65 127 _ _ PUNCT njb-49 65 128 _ _ PUNCT njb-49 65 129 _ _ PUNCT njb-49 65 130 _ _ PUNCT njb-49 65 131 _ _ PUNCT njb-49 65 132 _ _ PUNCT njb-49 65 133 _ _ PUNCT njb-49 65 134 _ _ PUNCT njb-49 65 135 _ _ PUNCT njb-49 65 136 _ _ PUNCT njb-49 65 137 _ _ PUNCT njb-49 65 138 _ _ PUNCT njb-49 65 139 _ _ PUNCT njb-49 65 140 _ _ PUNCT njb-49 65 141 _ _ PUNCT njb-49 65 142 _ _ PUNCT njb-49 65 143 _ _ PUNCT njb-49 65 144 _ _ PUNCT njb-49 65 145 _ _ PUNCT njb-49 65 146 _ _ PUNCT njb-49 65 147 _ _ PUNCT njb-49 65 148 _ _ PUNCT njb-49 65 149 _ _ PUNCT njb-49 65 150 _ _ PUNCT njb-49 65 151 _ _ PUNCT njb-49 65 152 _ _ PUNCT njb-49 65 153 _ _ PUNCT njb-49 65 154 _ _ PUNCT njb-49 65 155 _ _ PUNCT njb-49 65 156 _ _ PUNCT njb-49 65 157 _ _ PUNCT njb-49 65 158 _ _ PUNCT njb-49 65 159 _ _ PUNCT njb-49 65 160 _ _ PUNCT njb-49 65 161 _ _ PUNCT njb-49 65 162 _ _ PUNCT njb-49 65 163 _ _ PUNCT njb-49 65 164 _ _ PUNCT njb-49 65 165 _ _ PUNCT njb-49 65 166 _ _ PUNCT njb-49 65 167 _ _ PUNCT njb-49 65 168 _ _ PUNCT njb-49 65 169 _ _ PUNCT njb-49 65 170 _ _ PUNCT njb-49 65 171 _ _ PUNCT njb-49 65 172 _ _ PUNCT njb-49 65 173 _ _ PUNCT njb-49 65 174 _ _ PUNCT njb-49 65 175 _ _ PUNCT njb-49 65 176 _ _ PUNCT njb-49 65 177 _ _ PUNCT njb-49 65 178 _ _ PUNCT njb-49 65 179 _ _ PUNCT njb-49 65 180 _ _ PUNCT njb-49 65 181 _ _ PUNCT njb-49 65 182 _ _ PUNCT njb-49 65 183 _ _ PUNCT njb-49 65 184 _ _ PUNCT njb-49 65 185 _ _ PUNCT njb-49 66 1 fig.1 fig.1 ADJ njb-49 66 2 : : PUNCT njb-49 66 3 schematic schematic ADJ njb-49 66 4 diagram diagram PROPN njb-49 66 5 demonstrating demonstrating NOUN njb-49 66 6 bont bont PROPN njb-49 66 7 / / SYM njb-49 66 8 f f PROPN njb-49 66 9 domains domain NOUN njb-49 66 10 for for ADP njb-49 66 11 cloning clone VERB njb-49 66 12 . . PUNCT njb-49 67 1 nepal nepal ADJ njb-49 67 2     SPACE njb-49 67 3 journal journal NOUN njb-49 67 4     SPACE njb-49 67 5 of of ADP njb-49 67 6     SPACE njb-49 67 7 biotechnology biotechnology NOUN njb-49 67 8 . . PUNCT njb-49 67 9         SPACE njb-49 68 1 jan jan PROPN njb-49 68 2 . . PROPN njb-49 68 3     SPACE njb-49 68 4 2012 2012 NUM njb-49 68 5 , , PUNCT njb-49 68 6     SPACE njb-49 68 7 vol vol NOUN njb-49 68 8 . . PUNCT njb-49 68 9     SPACE njb-49 68 10 2 2 NUM njb-49 68 11 , , PUNCT njb-49 68 12     SPACE njb-49 68 13 no no INTJ njb-49 68 14 . . PUNCT njb-49 68 15     SPACE njb-49 69 1 1 1 NUM njb-49 69 2 : : PUNCT njb-49 69 3     SPACE njb-49 69 4 1 1 X njb-49 69 5     SPACE njb-49 69 6 – – PUNCT njb-49 69 7     SPACE njb-49 69 8 15 15 NUM njb-49 69 9                                                                                                                                                                                                                                         SPACE njb-49 69 10 biotechnology biotechnology NOUN njb-49 69 11     SPACE njb-49 69 12 society society NOUN njb-49 69 13     SPACE njb-49 69 14 of of ADP njb-49 69 15     SPACE njb-49 69 16 nepal nepal NOUN njb-49 69 17     SPACE njb-49 69 18 ( ( PUNCT njb-49 69 19 bsn bsn PROPN njb-49 69 20 ) ) PUNCT njb-49 69 21 , , PUNCT njb-49 69 22     SPACE njb-49 69 23 all all DET njb-49 69 24     SPACE njb-49 69 25 rights right NOUN njb-49 69 26     SPACE njb-49 69 27 reserved reserve VERB njb-49 69 28     SPACE njb-49 69 29 4 4 NUM njb-49 69 30 carried carry VERB njb-49 69 31 out out ADP njb-49 69 32 for for ADP njb-49 69 33 an an DET njb-49 69 34 additional additional ADJ njb-49 69 35 7 7 NUM njb-49 69 36 min min NOUN njb-49 69 37 at at ADP njb-49 69 38 72 72 NUM njb-49 69 39 ° ° NOUN njb-49 69 40 c c NOUN njb-49 69 41 . . PUNCT njb-49 70 1 amplified amplify VERB njb-49 70 2 pcr pcr PROPN njb-49 70 3 products product NOUN njb-49 70 4 were be AUX njb-49 70 5 purified purify VERB njb-49 70 6 by by ADP njb-49 70 7 agarose agarose NOUN njb-49 70 8 gel gel NOUN njb-49 70 9 elution elution NOUN njb-49 70 10 kit kit NOUN njb-49 70 11 ( ( PUNCT njb-49 70 12 intron intron PROPN njb-49 70 13 biotechnology biotechnology PROPN njb-49 70 14 , , PUNCT njb-49 70 15 the the DET njb-49 70 16 republic republic PROPN njb-49 70 17 of of ADP njb-49 70 18 korea korea PROPN njb-49 70 19 ) ) PUNCT njb-49 70 20 , , PUNCT njb-49 70 21 and and CCONJ njb-49 70 22 resultant resultant VERB njb-49 70 23 pcr pcr NOUN njb-49 70 24 products product NOUN njb-49 70 25 were be AUX njb-49 70 26 digested digest VERB njb-49 70 27 with with ADP njb-49 70 28 restriction restriction NOUN njb-49 70 29 enzymes enzyme NOUN njb-49 70 30 ( ( PUNCT njb-49 70 31 table table NOUN njb-49 70 32 1 1 NUM njb-49 70 33 ) ) PUNCT njb-49 70 34 and and CCONJ njb-49 70 35 subcloned subclone VERB njb-49 70 36 into into ADP njb-49 70 37 pgex4t-1 pgex4t-1 PROPN njb-49 70 38 vector vector NOUN njb-49 70 39 using use VERB njb-49 70 40 t4 t4 PROPN njb-49 70 41 ligase ligase PROPN njb-49 70 42 kit kit PROPN njb-49 70 43 ( ( PUNCT njb-49 70 44 promega promega PROPN njb-49 70 45 ) ) PUNCT njb-49 70 46 for for ADP njb-49 70 47 overnight overnight ADV njb-49 70 48 at at ADP njb-49 70 49 16 16 NUM njb-49 70 50 ° ° NOUN njb-49 70 51 c c NOUN njb-49 70 52 , , PUNCT njb-49 70 53 so so SCONJ njb-49 70 54 that that SCONJ njb-49 70 55 the the DET njb-49 70 56 correct correct ADJ njb-49 70 57 reading reading NOUN njb-49 70 58 frame frame NOUN njb-49 70 59 was be AUX njb-49 70 60 incorporated incorporate VERB njb-49 70 61 along along ADP njb-49 70 62 the the DET njb-49 70 63 thrombin thrombin NOUN njb-49 70 64 cleavage cleavage NOUN njb-49 70 65 site site NOUN njb-49 70 66 under under ADP njb-49 70 67 the the DET njb-49 70 68 gst gst NOUN njb-49 70 69 gene gene NOUN njb-49 70 70 ( ( PUNCT njb-49 70 71 representative representative ADJ njb-49 70 72 constructs construct NOUN njb-49 70 73 made make VERB njb-49 70 74 from from ADP njb-49 70 75 macvector macvector NOUN njb-49 70 76 10.0.2 10.0.2 NUM njb-49 70 77 ( ( PUNCT njb-49 70 78 nc nc PROPN njb-49 70 79 , , PUNCT njb-49 70 80 usa usa PROPN njb-49 70 81 ) ) PUNCT njb-49 70 82 software software NOUN njb-49 70 83 containing contain VERB njb-49 70 84 f f PROPN njb-49 70 85 - - PUNCT njb-49 70 86 lc lc NOUN njb-49 70 87 and and CCONJ njb-49 70 88 f f PROPN njb-49 70 89 - - PUNCT njb-49 70 90 hn hn PROPN njb-49 70 91 domains domain NOUN njb-49 70 92 are be AUX njb-49 70 93 shown show VERB njb-49 70 94 in in ADP njb-49 70 95 fig fig PROPN njb-49 70 96 . . PUNCT njb-49 70 97 2 2 NUM njb-49 70 98 ) ) PUNCT njb-49 70 99 . . PUNCT njb-49 71 1 ligated ligated ADJ njb-49 71 2 samples sample NOUN njb-49 71 3 were be AUX njb-49 71 4 transformed transform VERB njb-49 71 5 into into ADP njb-49 71 6 dh5α dh5α PROPN njb-49 71 7 by by ADP njb-49 71 8 heat heat NOUN njb-49 71 9 - - PUNCT njb-49 71 10 shock shock NOUN njb-49 71 11 ( ( PUNCT njb-49 71 12 42 42 NUM njb-49 71 13 ° ° NOUN njb-49 71 14 c c NOUN njb-49 71 15 for for ADP njb-49 71 16 50 50 NUM njb-49 71 17 sec sec PROPN njb-49 71 18 ) ) PUNCT njb-49 71 19 method method NOUN njb-49 71 20 and and CCONJ njb-49 71 21 positive positive ADJ njb-49 71 22 clones clone NOUN njb-49 71 23 with with ADP njb-49 71 24 appropriate appropriate ADJ njb-49 71 25 insert insert NOUN njb-49 71 26 were be AUX njb-49 71 27 screened screen VERB njb-49 71 28 out out ADP njb-49 71 29 by by ADP njb-49 71 30 use use NOUN njb-49 71 31 of of ADP njb-49 71 32 lb lb NOUN njb-49 71 33 agar agar NOUN njb-49 71 34 plates plate NOUN njb-49 71 35 containing contain VERB njb-49 71 36 100 100 NUM njb-49 71 37 µg µg PROPN njb-49 71 38 / / SYM njb-49 71 39 ml ml PROPN njb-49 71 40 ampicillin ampicillin PROPN njb-49 71 41 . . PUNCT njb-49 72 1 positive positive ADJ njb-49 72 2 colonies colony NOUN njb-49 72 3 containing contain VERB njb-49 72 4 the the DET njb-49 72 5 ligated ligate VERB njb-49 72 6 constructs construct NOUN njb-49 72 7 were be AUX njb-49 72 8 transformed transform VERB njb-49 72 9 into into ADP njb-49 72 10 bl21 bl21 NOUN njb-49 72 11 - - PUNCT njb-49 72 12 ril ril NOUN njb-49 72 13 for for ADP njb-49 72 14 the the DET njb-49 72 15 expression expression NOUN njb-49 72 16 of of ADP njb-49 72 17 type type NOUN njb-49 72 18 f f PROPN njb-49 72 19 recombinant recombinant ADJ njb-49 72 20 domains domain NOUN njb-49 72 21 . . PUNCT njb-49 73 1 expression expression NOUN njb-49 73 2 and and CCONJ njb-49 73 3 purification purification NOUN njb-49 73 4 of of ADP njb-49 73 5 recombinant recombinant ADJ njb-49 73 6 proteins protein NOUN njb-49 73 7 to to PART njb-49 73 8 monitor monitor VERB njb-49 73 9 the the DET njb-49 73 10 induction induction NOUN njb-49 73 11 of of ADP njb-49 73 12 recombinant recombinant ADJ njb-49 73 13 domains domain NOUN njb-49 73 14 in in ADP njb-49 73 15 bl21 bl21 NOUN njb-49 73 16 - - PUNCT njb-49 73 17 ril ril PROPN njb-49 73 18 , , PUNCT njb-49 73 19 2 2 NUM njb-49 73 20 ml ml NOUN njb-49 73 21 of of ADP njb-49 73 22 transformant transformant ADJ njb-49 73 23 cultures culture NOUN njb-49 73 24 were be AUX njb-49 73 25 inoculated inoculate VERB njb-49 73 26 and and CCONJ njb-49 73 27 grown grow VERB njb-49 73 28 in in ADP njb-49 73 29 lb lb NOUN njb-49 73 30 broth broth NOUN njb-49 73 31 and and CCONJ njb-49 73 32 induced induce VERB njb-49 73 33 at at ADP njb-49 73 34 various various ADJ njb-49 73 35 time time NOUN njb-49 73 36 intervals interval NOUN njb-49 73 37 using use VERB njb-49 73 38 different different ADJ njb-49 73 39 concentrations concentration NOUN njb-49 73 40 of of ADP njb-49 73 41 isopropyl isopropyl PROPN njb-49 73 42 β β NOUN njb-49 73 43 - - PUNCT njb-49 73 44 d-1 d-1 ADJ njb-49 73 45 - - PUNCT njb-49 73 46 thiogalactopyranoside thiogalactopyranoside NOUN njb-49 73 47 ( ( PUNCT njb-49 73 48 iptg iptg PROPN njb-49 73 49 ) ) PUNCT njb-49 73 50 . . PUNCT njb-49 74 1 in in ADP njb-49 74 2 order order NOUN njb-49 74 3 to to PART njb-49 74 4 purify purify VERB njb-49 74 5 recombinant recombinant ADJ njb-49 74 6 proteins protein NOUN njb-49 74 7 , , PUNCT njb-49 74 8 mass mass ADJ njb-49 74 9 production production NOUN njb-49 74 10 was be AUX njb-49 74 11 performed perform VERB njb-49 74 12 by by ADP njb-49 74 13 inoculating inoculate VERB njb-49 74 14 2 2 NUM njb-49 74 15 ml ml NOUN njb-49 74 16 induced induce VERB njb-49 74 17 culture culture NOUN njb-49 74 18 in in ADP njb-49 74 19 1 1 NUM njb-49 74 20 liter liter NOUN njb-49 74 21 followed follow VERB njb-49 74 22 by by ADP njb-49 74 23 centrifugation centrifugation NOUN njb-49 74 24 ( ( PUNCT njb-49 74 25 12,000 12,000 NUM njb-49 74 26 xg xg NUM njb-49 74 27 ) ) PUNCT njb-49 74 28 at at ADP njb-49 74 29 4 4 NUM njb-49 74 30 ° ° NOUN njb-49 74 31 c c NOUN njb-49 74 32 for for ADP njb-49 74 33 20 20 NUM njb-49 74 34 min min PROPN njb-49 74 35 . . PROPN njb-49 74 36 pellet pellet PROPN njb-49 74 37 was be AUX njb-49 74 38 then then ADV njb-49 74 39 resuspended resuspend VERB njb-49 74 40 in in ADP njb-49 74 41 10 10 NUM njb-49 74 42 ml ml NOUN njb-49 74 43 of of ADP njb-49 74 44 50 50 NUM njb-49 74 45 mm mm NOUN njb-49 74 46 phosphate phosphate NOUN njb-49 74 47 buffered buffer VERB njb-49 74 48 saline saline NOUN njb-49 74 49 ( ( PUNCT njb-49 74 50 pbs pbs PROPN njb-49 74 51 ) ) PUNCT njb-49 74 52 and and CCONJ njb-49 74 53 cells cell NOUN njb-49 74 54 were be AUX njb-49 74 55 then then ADV njb-49 74 56 lysed lyse VERB njb-49 74 57 by by ADP njb-49 74 58 sonication sonication NOUN njb-49 74 59 , , PUNCT njb-49 74 60 with with SCONJ njb-49 74 61 10 10 NUM njb-49 74 62 short short ADJ njb-49 74 63 bursts burst NOUN njb-49 74 64 of of ADP njb-49 74 65 30 30 NUM njb-49 74 66 sec sec PROPN njb-49 74 67 followed follow VERB njb-49 74 68 by by ADP njb-49 74 69 intervals interval NOUN njb-49 74 70 of of ADP njb-49 74 71 placing place VERB njb-49 74 72 samples sample NOUN njb-49 74 73 on on ADP njb-49 74 74 ice ice NOUN njb-49 74 75 for for ADP njb-49 74 76 1 1 NUM njb-49 74 77 min min NOUN njb-49 74 78 cooling cooling NOUN njb-49 74 79 . . PUNCT njb-49 75 1 2 2 NUM njb-49 75 2 ( ( PUNCT njb-49 75 3 a a PRON njb-49 75 4 ) ) PUNCT njb-49 75 5 2 2 NUM njb-49 75 6 ( ( PUNCT njb-49 75 7 b b NOUN njb-49 75 8 ) ) PUNCT njb-49 75 9 fig.2 fig.2 X njb-49 75 10 : : PUNCT njb-49 75 11 schematic schematic ADJ njb-49 75 12 diagram diagram PROPN njb-49 75 13 of of ADP njb-49 75 14 constructed construct VERB njb-49 75 15 plasmids plasmid NOUN njb-49 75 16 generated generate VERB njb-49 75 17 by by ADP njb-49 75 18 macvector macvector NOUN njb-49 75 19 10.0.2 10.0.2 NUM njb-49 75 20 software software NOUN njb-49 75 21 ( ( PUNCT njb-49 75 22 symantec symantec ADJ njb-49 75 23 corporation corporation NOUN njb-49 75 24 ) ) PUNCT njb-49 75 25 . . PUNCT njb-49 76 1 plasmids plasmid NOUN njb-49 76 2 pgex-4t-1 pgex-4t-1 PROPN njb-49 76 3 - - PUNCT njb-49 76 4 f f PROPN njb-49 76 5 - - PUNCT njb-49 76 6 lc lc PROPN njb-49 76 7 ( ( PUNCT njb-49 76 8 a a NOUN njb-49 76 9 ) ) PUNCT njb-49 76 10 and and CCONJ njb-49 76 11 pgex-4t-1 pgex-4t-1 PROPN njb-49 76 12 - - PUNCT njb-49 76 13 f f PROPN njb-49 76 14 - - PUNCT njb-49 76 15 hn hn PROPN njb-49 76 16 ( ( PUNCT njb-49 76 17 b b PROPN njb-49 76 18 ) ) PUNCT njb-49 76 19 encoding encode VERB njb-49 76 20 catalytic catalytic ADJ njb-49 76 21 and and CCONJ njb-49 76 22 translocation translocation NOUN njb-49 76 23 domains domain NOUN njb-49 76 24 . . PUNCT njb-49 77 1 nepal nepal ADJ njb-49 77 2     SPACE njb-49 77 3 journal journal NOUN njb-49 77 4     SPACE njb-49 77 5 of of ADP njb-49 77 6     SPACE njb-49 77 7 biotechnology biotechnology NOUN njb-49 77 8 . . PUNCT njb-49 77 9         SPACE njb-49 78 1 jan jan PROPN njb-49 78 2 . . PROPN njb-49 78 3     SPACE njb-49 78 4 2012 2012 NUM njb-49 78 5 , , PUNCT njb-49 78 6     SPACE njb-49 78 7 vol vol NOUN njb-49 78 8 . . PUNCT njb-49 78 9     SPACE njb-49 78 10 2 2 NUM njb-49 78 11 , , PUNCT njb-49 78 12     SPACE njb-49 78 13 no no INTJ njb-49 78 14 . . PUNCT njb-49 78 15     SPACE njb-49 79 1 1 1 NUM njb-49 79 2 : : PUNCT njb-49 79 3     SPACE njb-49 79 4 1 1 X njb-49 79 5     SPACE njb-49 79 6 – – PUNCT njb-49 79 7     SPACE njb-49 79 8 15 15 NUM njb-49 79 9                                                                                                                                                                                                                                         SPACE njb-49 79 10 biotechnology biotechnology NOUN njb-49 79 11     SPACE njb-49 79 12 society society NOUN njb-49 79 13     SPACE njb-49 79 14 of of ADP njb-49 79 15     SPACE njb-49 79 16 nepal nepal NOUN njb-49 79 17     SPACE njb-49 79 18 ( ( PUNCT njb-49 79 19 bsn bsn PROPN njb-49 79 20 ) ) PUNCT njb-49 79 21 , , PUNCT njb-49 79 22     SPACE njb-49 79 23 all all DET njb-49 79 24     SPACE njb-49 79 25 rights right NOUN njb-49 79 26     SPACE njb-49 79 27 reserved reserve VERB njb-49 79 28     SPACE njb-49 79 29 5 5 NUM njb-49 79 30 after after ADP njb-49 79 31 centrifugation centrifugation NOUN njb-49 79 32 at at ADP njb-49 79 33 12,000 12,000 NUM njb-49 79 34 xg xg NOUN njb-49 79 35 for for ADP njb-49 79 36 30 30 NUM njb-49 79 37 min min NOUN njb-49 79 38 , , PUNCT njb-49 79 39 the the DET njb-49 79 40 pellet pellet NOUN njb-49 79 41 was be AUX njb-49 79 42 resuspended resuspend VERB njb-49 79 43 in in ADP njb-49 79 44 10 10 NUM njb-49 79 45 ml ml NOUN njb-49 79 46 pbs pbs NOUN njb-49 79 47 overnight overnight ADV njb-49 79 48 at at SCONJ njb-49 79 49 4 4 NUM njb-49 79 50 ° ° NOUN njb-49 79 51 c c NOUN njb-49 79 52 and and CCONJ njb-49 79 53 supernatant supernatant NOUN njb-49 79 54 was be AUX njb-49 79 55 placed place VERB njb-49 79 56 in in ADP njb-49 79 57 a a DET njb-49 79 58 gst gst NOUN njb-49 79 59 affinity affinity NOUN njb-49 79 60 chromatography chromatography NOUN njb-49 79 61 column column NOUN njb-49 79 62 after after SCONJ njb-49 79 63 column column NOUN njb-49 79 64 was be AUX njb-49 79 65 initially initially ADV njb-49 79 66 washed wash VERB njb-49 79 67 multiple multiple ADJ njb-49 79 68 times time NOUN njb-49 79 69 with with ADP njb-49 79 70 deionized deionized ADJ njb-49 79 71 water water NOUN njb-49 79 72 and and CCONJ njb-49 79 73 pbs pbs PROPN njb-49 79 74 , , PUNCT njb-49 79 75 followed follow VERB njb-49 79 76 by by ADP njb-49 79 77 normalization normalization NOUN njb-49 79 78 with with ADP njb-49 79 79 gst gst NOUN njb-49 79 80 resin resin NOUN njb-49 79 81 . . PUNCT njb-49 80 1 the the DET njb-49 80 2 large large ADJ njb-49 80 3 scale scale NOUN njb-49 80 4 of of ADP njb-49 80 5 gst gst NOUN njb-49 80 6 - - PUNCT njb-49 80 7 fusion fusion NOUN njb-49 80 8 proteins protein NOUN njb-49 80 9 was be AUX njb-49 80 10 eluted elute VERB njb-49 80 11 by by ADP njb-49 80 12 single single ADJ njb-49 80 13 step step NOUN njb-49 80 14 affinity affinity NOUN njb-49 80 15 chromatography chromatography NOUN njb-49 80 16 ( ( PUNCT njb-49 80 17 containing contain VERB njb-49 80 18 sepharose sepharose NOUN njb-49 80 19 4b 4b PROPN njb-49 80 20 beads bead NOUN njb-49 80 21 ) ) PUNCT njb-49 80 22 . . PUNCT njb-49 81 1 fractions fraction NOUN njb-49 81 2 containing contain VERB njb-49 81 3 desired desire VERB njb-49 81 4 proteins protein NOUN njb-49 81 5 were be AUX njb-49 81 6 pooled pool VERB njb-49 81 7 and and CCONJ njb-49 81 8 dialyzed dialyze VERB njb-49 81 9 for for ADP njb-49 81 10 2 2 NUM njb-49 81 11 h h NOUN njb-49 81 12 at at ADP njb-49 81 13 4 4 NUM njb-49 81 14 ° ° NOUN njb-49 81 15 c c NOUN njb-49 81 16 against against ADP njb-49 81 17 pbs pbs PROPN njb-49 81 18 . . PUNCT njb-49 82 1 for for ADP njb-49 82 2 thrombin thrombin NOUN njb-49 82 3 protease protease NOUN njb-49 82 4 treatment treatment NOUN njb-49 82 5 , , PUNCT njb-49 82 6 resin resin NOUN njb-49 82 7 bound bind VERB njb-49 82 8 gst gst NOUN njb-49 82 9 - - PUNCT njb-49 82 10 fusion fusion NOUN njb-49 82 11 proteins protein NOUN njb-49 82 12 were be AUX njb-49 82 13 cleaved cleave VERB njb-49 82 14 with with ADP njb-49 82 15 20 20 NUM njb-49 82 16 units unit NOUN njb-49 82 17 thrombin thrombin NOUN njb-49 82 18 incubating incubate VERB njb-49 82 19 for for ADP njb-49 82 20 30 30 NUM njb-49 82 21 min min NOUN njb-49 82 22 at at ADP njb-49 82 23 37 37 NUM njb-49 82 24 ° ° NOUN njb-49 82 25 c c NOUN njb-49 82 26 followed follow VERB njb-49 82 27 by by ADP njb-49 82 28 15 15 NUM njb-49 82 29 min min NOUN njb-49 82 30 at at ADP njb-49 82 31 room room NOUN njb-49 82 32 temp temp NOUN njb-49 82 33 ( ( PUNCT njb-49 82 34 20 20 NUM njb-49 82 35 25 25 NUM njb-49 82 36 ° ° NUM njb-49 82 37 c c NOUN njb-49 82 38 ) ) PUNCT njb-49 82 39 and and CCONJ njb-49 82 40 final final ADJ njb-49 82 41 elution elution NOUN njb-49 82 42 with with ADP njb-49 82 43 500 500 NUM njb-49 82 44 µl µl PROPN njb-49 82 45 pbs pbs NOUN njb-49 82 46 . . PUNCT njb-49 83 1 the the DET njb-49 83 2 eluted elute VERB njb-49 83 3 proteins protein NOUN njb-49 83 4 were be AUX njb-49 83 5 further far ADV njb-49 83 6 dialyzed dialyze VERB njb-49 83 7 to to PART njb-49 83 8 maintain maintain VERB njb-49 83 9 salt salt NOUN njb-49 83 10 concentrations concentration NOUN njb-49 83 11 and and CCONJ njb-49 83 12 ph ph NOUN njb-49 83 13 . . PUNCT njb-49 84 1 protein protein NOUN njb-49 84 2 concentrations concentration NOUN njb-49 84 3 were be AUX njb-49 84 4 measured measure VERB njb-49 84 5 according accord VERB njb-49 84 6 to to ADP njb-49 84 7 the the DET njb-49 84 8 bradford bradford PROPN njb-49 84 9 method method PROPN njb-49 84 10 using use VERB njb-49 84 11 a a DET njb-49 84 12 bio bio ADJ njb-49 84 13 - - ADJ njb-49 84 14 rad rad ADJ njb-49 84 15 model model NOUN njb-49 84 16 550 550 NUM njb-49 84 17 microtiter microtiter NOUN njb-49 84 18 plate plate NOUN njb-49 84 19 reader reader NOUN njb-49 84 20 . . PUNCT njb-49 85 1 all all DET njb-49 85 2 proteins protein NOUN njb-49 85 3 were be AUX njb-49 85 4 labeled label VERB njb-49 85 5 , , PUNCT njb-49 85 6 aliquoted aliquote VERB njb-49 85 7 and and CCONJ njb-49 85 8 stored store VERB njb-49 85 9 at at ADP njb-49 85 10 – – PUNCT njb-49 85 11 70 70 NUM njb-49 85 12 ° ° NOUN njb-49 85 13 c c NOUN njb-49 85 14 prior prior ADV njb-49 85 15 to to ADP njb-49 85 16 use use VERB njb-49 85 17 . . PUNCT njb-49 86 1 lastly lastly ADV njb-49 86 2 , , PUNCT njb-49 86 3 sodium sodium PROPN njb-49 86 4 dodecyl dodecyl PROPN njb-49 86 5 sulfate sulfate NOUN njb-49 86 6 polyacrylamide polyacrylamide NOUN njb-49 86 7 gel gel NOUN njb-49 86 8 electrophoresis electrophoresis NOUN njb-49 86 9 ( ( PUNCT njb-49 86 10 sds sds NOUN njb-49 86 11 - - PUNCT njb-49 86 12 page page NOUN njb-49 86 13 ) ) PUNCT njb-49 86 14 was be AUX njb-49 86 15 carried carry VERB njb-49 86 16 out out ADP njb-49 86 17 on on ADP njb-49 86 18 a a DET njb-49 86 19 12 12 NUM njb-49 86 20 % % NOUN njb-49 86 21 gel gel NOUN njb-49 86 22 under under ADP njb-49 86 23 reducing reduce VERB njb-49 86 24 conditions condition NOUN njb-49 86 25 in in ADP njb-49 86 26 order order NOUN njb-49 86 27 to to PART njb-49 86 28 determine determine VERB njb-49 86 29 solubility solubility NOUN njb-49 86 30 of of ADP njb-49 86 31 the the DET njb-49 86 32 eluted elute VERB njb-49 86 33 proteins protein NOUN njb-49 86 34 . . PUNCT njb-49 87 1 production production NOUN njb-49 87 2 and and CCONJ njb-49 87 3 analysis analysis NOUN njb-49 87 4 of of ADP njb-49 87 5 polyclonal polyclonal ADJ njb-49 87 6 antibodies antibody NOUN njb-49 87 7 against against ADP njb-49 87 8 rf rf NOUN njb-49 87 9 - - PUNCT njb-49 87 10 lc lc NOUN njb-49 87 11 and and CCONJ njb-49 87 12 rf rf NOUN njb-49 87 13 - - NOUN njb-49 87 14 hn hn NOUN njb-49 87 15 for for ADP njb-49 87 16 immunization immunization NOUN njb-49 87 17 , , PUNCT njb-49 87 18 three three NUM njb-49 87 19 female female ADJ njb-49 87 20 balb balb NOUN njb-49 87 21 / / SYM njb-49 87 22 c c NOUN njb-49 87 23 mice mouse NOUN njb-49 87 24 ( ( PUNCT njb-49 87 25 four four NUM njb-49 87 26 weeks week NOUN njb-49 87 27 old old ADJ njb-49 87 28 having have VERB njb-49 87 29 around around ADP njb-49 87 30 18 18 NUM njb-49 87 31 - - SYM njb-49 87 32 22 22 NUM njb-49 87 33 gram gram NOUN njb-49 87 34 body body NOUN njb-49 87 35 weight weight NOUN njb-49 87 36 ) ) PUNCT njb-49 87 37 were be AUX njb-49 87 38 immunized immunize VERB njb-49 87 39 intraperitoneally intraperitoneally ADV njb-49 87 40 with with ADP njb-49 87 41 10 10 NUM njb-49 87 42 µg µg NOUN njb-49 87 43 of of ADP njb-49 87 44 antigen antigen NOUN njb-49 87 45 mixtures mixture NOUN njb-49 87 46 containing contain VERB njb-49 87 47 recombinant recombinant ADJ njb-49 87 48 proteins protein NOUN njb-49 87 49 ( ( PUNCT njb-49 87 50 25 25 NUM njb-49 87 51 µg µg NOUN njb-49 87 52 gst gst X njb-49 87 53 free free ADJ njb-49 87 54 f f X njb-49 87 55 - - PUNCT njb-49 87 56 lc lc NOUN njb-49 87 57 or or CCONJ njb-49 87 58 25 25 NUM njb-49 87 59 µg µg NOUN njb-49 87 60 gst gst X njb-49 87 61 free free ADJ njb-49 87 62 f f X njb-49 87 63 - - PUNCT njb-49 87 64 hn hn PROPN njb-49 87 65 ) ) PUNCT njb-49 87 66 in in ADP njb-49 87 67 pbs pbs PROPN njb-49 87 68 . . PUNCT njb-49 88 1 additionally additionally ADV njb-49 88 2 , , PUNCT njb-49 88 3 an an DET njb-49 88 4 equal equal ADJ njb-49 88 5 volume volume NOUN njb-49 88 6 of of ADP njb-49 88 7 freund freund PROPN njb-49 88 8 ’s ’s PART njb-49 88 9 complete complete ADJ njb-49 88 10 adjuvant adjuvant NOUN njb-49 88 11 was be AUX njb-49 88 12 separately separately ADV njb-49 88 13 injected inject VERB njb-49 88 14 into into ADP njb-49 88 15 each each DET njb-49 88 16 mouse mouse NOUN njb-49 88 17 . . PUNCT njb-49 89 1 after after ADP njb-49 89 2 two two NUM njb-49 89 3 weeks week NOUN njb-49 89 4 lapse lapse NOUN njb-49 89 5 , , PUNCT njb-49 89 6 the the DET njb-49 89 7 mice mouse NOUN njb-49 89 8 were be AUX njb-49 89 9 immunized immunize VERB njb-49 89 10 with with ADP njb-49 89 11 the the DET njb-49 89 12 mixture mixture NOUN njb-49 89 13 containing contain VERB njb-49 89 14 20 20 NUM njb-49 89 15 µg µg NOUN njb-49 89 16 of of ADP njb-49 89 17 recombinant recombinant ADJ njb-49 89 18 protein protein NOUN njb-49 89 19 and and CCONJ njb-49 89 20 equal equal ADJ njb-49 89 21 volume volume NOUN njb-49 89 22 of of ADP njb-49 89 23 freund freund PROPN njb-49 89 24 ’s ’s PART njb-49 89 25 incomplete incomplete ADJ njb-49 89 26 adjuvant adjuvant NOUN njb-49 89 27 . . PUNCT njb-49 90 1 at at ADP njb-49 90 2 the the DET njb-49 90 3 start start NOUN njb-49 90 4 of of ADP njb-49 90 5 the the DET njb-49 90 6 fourth fourth ADJ njb-49 90 7 week week NOUN njb-49 90 8 following follow VERB njb-49 90 9 the the DET njb-49 90 10 initial initial ADJ njb-49 90 11 immunization immunization NOUN njb-49 90 12 , , PUNCT njb-49 90 13 the the DET njb-49 90 14 mice mouse NOUN njb-49 90 15 were be AUX njb-49 90 16 finally finally ADV njb-49 90 17 boosted boost VERB njb-49 90 18 by by ADP njb-49 90 19 intravenous intravenous ADJ njb-49 90 20 injection injection NOUN njb-49 90 21 of of ADP njb-49 90 22 the the DET njb-49 90 23 recombinant recombinant ADJ njb-49 90 24 proteins protein NOUN njb-49 90 25 ( ( PUNCT njb-49 90 26 30 30 NUM njb-49 90 27 µg µg NOUN njb-49 90 28 ) ) PUNCT njb-49 90 29 without without ADP njb-49 90 30 adjuvant adjuvant NOUN njb-49 90 31 . . PUNCT njb-49 91 1 three three NUM njb-49 91 2 days day NOUN njb-49 91 3 following follow VERB njb-49 91 4 final final ADJ njb-49 91 5 boost boost NOUN njb-49 91 6 , , PUNCT njb-49 91 7 mice mouse NOUN njb-49 91 8 were be AUX njb-49 91 9 then then ADV njb-49 91 10 bled bleed VERB njb-49 91 11 from from ADP njb-49 91 12 the the DET njb-49 91 13 tail tail NOUN njb-49 91 14 and and CCONJ njb-49 91 15 tested test VERB njb-49 91 16 by by ADP njb-49 91 17 indirect indirect ADJ njb-49 91 18 elisa elisa NOUN njb-49 91 19 . . PUNCT njb-49 92 1 preparation preparation NOUN njb-49 92 2 of of ADP njb-49 92 3 monoclonal monoclonal NOUN njb-49 92 4 antibodies antibody NOUN njb-49 92 5 ( ( PUNCT njb-49 92 6 hybridoma hybridoma NOUN njb-49 92 7 technology technology NOUN njb-49 92 8 ) ) PUNCT njb-49 92 9 cell cell NOUN njb-49 92 10 fusion fusion NOUN njb-49 92 11 and and CCONJ njb-49 92 12 culture culture NOUN njb-49 92 13 of of ADP njb-49 92 14 hybridomas hybridomas PROPN njb-49 92 15 was be AUX njb-49 92 16 carried carry VERB njb-49 92 17 out out ADP njb-49 92 18 according accord VERB njb-49 92 19 to to ADP njb-49 92 20 the the DET njb-49 92 21 protocol protocol NOUN njb-49 92 22 by by ADP njb-49 92 23 kohler kohler NOUN njb-49 92 24 et et PROPN njb-49 92 25 al al PROPN njb-49 92 26 . . PUNCT njb-49 93 1 [ [ X njb-49 93 2 24 24 NUM njb-49 93 3 ] ] PUNCT njb-49 93 4 . . PUNCT njb-49 94 1 briefly briefly ADV njb-49 94 2 , , PUNCT njb-49 94 3 cultures culture NOUN njb-49 94 4 were be AUX njb-49 94 5 incubated incubate VERB njb-49 94 6 at at ADP njb-49 94 7 37 37 NUM njb-49 94 8 ° ° NOUN njb-49 94 9 c c NOUN njb-49 94 10 in in ADP njb-49 94 11 an an DET njb-49 94 12 incubator incubator NOUN njb-49 94 13 with with ADP njb-49 94 14 5 5 NUM njb-49 94 15 % % NOUN njb-49 94 16 co2 co2 NOUN njb-49 94 17 - - PUNCT njb-49 94 18 in in ADP njb-49 94 19 - - PUNCT njb-49 94 20 air air NOUN njb-49 94 21 and and CCONJ njb-49 94 22 98 98 NUM njb-49 94 23 % % NOUN njb-49 94 24 relative relative ADJ njb-49 94 25 humidity humidity NOUN njb-49 94 26 . . PUNCT njb-49 95 1 four four NUM njb-49 95 2 days day NOUN njb-49 95 3 following follow VERB njb-49 95 4 final final ADJ njb-49 95 5 boost boost NOUN njb-49 95 6 , , PUNCT njb-49 95 7 the the DET njb-49 95 8 mouse mouse NOUN njb-49 95 9 showing show VERB njb-49 95 10 the the DET njb-49 95 11 highest high ADJ njb-49 95 12 antibody antibody NOUN njb-49 95 13 titer titer NOUN njb-49 95 14 by by ADP njb-49 95 15 elisa elisa NOUN njb-49 95 16 was be AUX njb-49 95 17 sacrificed sacrifice VERB njb-49 95 18 by by ADP njb-49 95 19 cervical cervical ADJ njb-49 95 20 dislocation dislocation NOUN njb-49 95 21 and and CCONJ njb-49 95 22 its its PRON njb-49 95 23 spleen spleen NOUN njb-49 95 24 was be AUX njb-49 95 25 removed remove VERB njb-49 95 26 aseptically aseptically ADV njb-49 95 27 . . PUNCT njb-49 96 1 spleen spleen PROPN njb-49 96 2 cells cell NOUN njb-49 96 3 ( ( PUNCT njb-49 96 4 1x108 1x108 NUM njb-49 96 5 viable viable ADJ njb-49 96 6 cells cell NOUN njb-49 96 7 ) ) PUNCT njb-49 96 8 were be AUX njb-49 96 9 mixed mix VERB njb-49 96 10 with with ADP njb-49 96 11 sp2/0ag sp2/0ag ADJ njb-49 96 12 14 14 NUM njb-49 96 13 myeloma myeloma NOUN njb-49 96 14 cells cell NOUN njb-49 96 15 ( ( PUNCT njb-49 96 16 1x107viable 1x107viable NUM njb-49 96 17 cells cell NOUN njb-49 96 18 ) ) PUNCT njb-49 96 19 and and CCONJ njb-49 96 20 nepal nepal ADJ njb-49 96 21     SPACE njb-49 96 22 journal journal NOUN njb-49 96 23     SPACE njb-49 96 24 of of ADP njb-49 96 25     SPACE njb-49 96 26 biotechnology biotechnology NOUN njb-49 96 27 . . PUNCT njb-49 96 28         SPACE njb-49 97 1 jan jan PROPN njb-49 97 2 . . PROPN njb-49 97 3     SPACE njb-49 97 4 2012 2012 NUM njb-49 97 5 , , PUNCT njb-49 97 6     SPACE njb-49 97 7 vol vol NOUN njb-49 97 8 . . PUNCT njb-49 97 9     SPACE njb-49 97 10 2 2 NUM njb-49 97 11 , , PUNCT njb-49 97 12     SPACE njb-49 97 13 no no INTJ njb-49 97 14 . . PUNCT njb-49 97 15     SPACE njb-49 98 1 1 1 NUM njb-49 98 2 : : PUNCT njb-49 98 3     SPACE njb-49 98 4 1 1 X njb-49 98 5     SPACE njb-49 98 6 – – PUNCT njb-49 98 7     SPACE njb-49 98 8 15 15 NUM njb-49 98 9                                                                                                                                                                                                                                         SPACE njb-49 98 10 biotechnology biotechnology NOUN njb-49 98 11     SPACE njb-49 98 12 society society NOUN njb-49 98 13     SPACE njb-49 98 14 of of ADP njb-49 98 15     SPACE njb-49 98 16 nepal nepal NOUN njb-49 98 17     SPACE njb-49 98 18 ( ( PUNCT njb-49 98 19 bsn bsn PROPN njb-49 98 20 ) ) PUNCT njb-49 98 21 , , PUNCT njb-49 98 22     SPACE njb-49 98 23 all all DET njb-49 98 24     SPACE njb-49 98 25 rights right NOUN njb-49 98 26     SPACE njb-49 98 27 reserved reserve VERB njb-49 98 28     SPACE njb-49 98 29 6 6 NUM njb-49 98 30 grown grow VERB njb-49 98 31 in in ADP njb-49 98 32 dulbecco dulbecco NOUN njb-49 98 33 ’s ’s PART njb-49 98 34 modified modify VERB njb-49 98 35 eagle eagle NOUN njb-49 98 36 ’s ’s PART njb-49 98 37 medium medium NOUN njb-49 98 38 ( ( PUNCT njb-49 98 39 dmem dmem PROPN njb-49 98 40 ) ) PUNCT njb-49 98 41 supplemented supplement VERB njb-49 98 42 with with ADP njb-49 98 43 10 10 NUM njb-49 98 44 % % NOUN njb-49 98 45 fetal fetal ADJ njb-49 98 46 bovine bovine NOUN njb-49 98 47 serum serum NOUN njb-49 98 48 ( ( PUNCT njb-49 98 49 fbs fbs ADJ njb-49 98 50 ) ) PUNCT njb-49 98 51 ( ( PUNCT njb-49 98 52 hyclone hyclone NOUN njb-49 98 53 ) ) PUNCT njb-49 98 54 . . PUNCT njb-49 99 1 the the DET njb-49 99 2 addition addition NOUN njb-49 99 3 of of ADP njb-49 99 4 50 50 NUM njb-49 99 5 % % NOUN njb-49 99 6 polyethylene polyethylene NOUN njb-49 99 7 glycol glycol PROPN njb-49 99 8 4000 4000 NUM njb-49 99 9 ( ( PUNCT njb-49 99 10 peg peg NOUN njb-49 99 11 ) ) PUNCT njb-49 99 12 ( ( PUNCT njb-49 99 13 sigma sigma PROPN njb-49 99 14 aldrich aldrich PROPN njb-49 99 15 ) ) PUNCT njb-49 99 16 allowed allow VERB njb-49 99 17 for for ADP njb-49 99 18 the the DET njb-49 99 19 fusion fusion NOUN njb-49 99 20 of of ADP njb-49 99 21 mixed mixed ADJ njb-49 99 22 cells cell NOUN njb-49 99 23 ; ; PUNCT njb-49 99 24 resulting result VERB njb-49 99 25 in in ADP njb-49 99 26 hybrid hybrid ADJ njb-49 99 27 cells cell NOUN njb-49 99 28 . . PUNCT njb-49 100 1 hybrid hybrid ADJ njb-49 100 2 cells cell NOUN njb-49 100 3 were be AUX njb-49 100 4 screened screen VERB njb-49 100 5 by by ADP njb-49 100 6 incubation incubation NOUN njb-49 100 7 in in ADP njb-49 100 8 96 96 NUM njb-49 100 9 - - PUNCT njb-49 100 10 well well NOUN njb-49 100 11 plates plate NOUN njb-49 100 12 ( ( PUNCT njb-49 100 13 nunc nunc NOUN njb-49 100 14 ) ) PUNCT njb-49 100 15 containing contain VERB njb-49 100 16 hypoxanthine hypoxanthine NOUN njb-49 100 17 - - PUNCT njb-49 100 18 aminopterin aminopterin NOUN njb-49 100 19 - - PUNCT njb-49 100 20 thymidine thymidine NOUN njb-49 100 21 ( ( PUNCT njb-49 100 22 hat hat NOUN njb-49 100 23 ) ) PUNCT njb-49 100 24 selection selection NOUN njb-49 100 25 medium medium NOUN njb-49 100 26 in in ADP njb-49 100 27 combination combination NOUN njb-49 100 28 with with ADP njb-49 100 29 mouse mouse NOUN njb-49 100 30 feeder feeder NOUN njb-49 100 31 cells cell NOUN njb-49 100 32 for for ADP njb-49 100 33 one one NUM njb-49 100 34 week week NOUN njb-49 100 35 . . PUNCT njb-49 101 1 antibody antibody NOUN njb-49 101 2 production production NOUN njb-49 101 3 from from ADP njb-49 101 4 hybrid hybrid ADJ njb-49 101 5 cells cell NOUN njb-49 101 6 was be AUX njb-49 101 7 monitored monitor VERB njb-49 101 8 through through ADP njb-49 101 9 indirect indirect ADJ njb-49 101 10 elisa elisa NOUN njb-49 101 11 ; ; PUNCT njb-49 101 12 positive positive ADJ njb-49 101 13 cells cell NOUN njb-49 101 14 were be AUX njb-49 101 15 expanded expand VERB njb-49 101 16 into into ADP njb-49 101 17 24 24 NUM njb-49 101 18 - - PUNCT njb-49 101 19 well well NOUN njb-49 101 20 plates plate NOUN njb-49 101 21 ( ( PUNCT njb-49 101 22 nunc nunc NOUN njb-49 101 23 ) ) PUNCT njb-49 101 24 containing contain VERB njb-49 101 25 hypoxanthine hypoxanthine NOUN njb-49 101 26 - - PUNCT njb-49 101 27 thymidine thymidine NOUN njb-49 101 28 ( ( PUNCT njb-49 101 29 ht ht NOUN njb-49 101 30 ) ) PUNCT njb-49 101 31 media medium NOUN njb-49 101 32 supplemented supplement VERB njb-49 101 33 with with ADP njb-49 101 34 dilute dilute PROPN njb-49 101 35 aminopterin aminopterin PROPN njb-49 101 36 according accord VERB njb-49 101 37 to to ADP njb-49 101 38 the the DET njb-49 101 39 standard standard ADJ njb-49 101 40 protocol protocol NOUN njb-49 101 41 by by ADP njb-49 101 42 harlow harlow PROPN njb-49 101 43 and and CCONJ njb-49 101 44 lane lane NOUN njb-49 101 45 [ [ PUNCT njb-49 101 46 25 25 NUM njb-49 101 47 ] ] PUNCT njb-49 101 48 . . PUNCT njb-49 102 1 elisa elisa PROPN njb-49 102 2 - - PUNCT njb-49 102 3 positive positive ADJ njb-49 102 4 hybridomas hybridoma NOUN njb-49 102 5 were be AUX njb-49 102 6 selected select VERB njb-49 102 7 and and CCONJ njb-49 102 8 cloned clone VERB njb-49 102 9 twice twice ADV njb-49 102 10 via via ADP njb-49 102 11 limiting limit VERB njb-49 102 12 dilution dilution NOUN njb-49 102 13 ; ; PUNCT njb-49 102 14 one one NUM njb-49 102 15 cell cell NOUN njb-49 102 16 per per ADP njb-49 102 17 well well ADV njb-49 102 18 into into ADP njb-49 102 19 96 96 NUM njb-49 102 20 - - PUNCT njb-49 102 21 well well NOUN njb-49 102 22 plates plate NOUN njb-49 102 23 supplemented supplement VERB njb-49 102 24 with with ADP njb-49 102 25 ht ht NOUN njb-49 102 26 media medium NOUN njb-49 102 27 and and CCONJ njb-49 102 28 feeder feeder NOUN njb-49 102 29 cells cell NOUN njb-49 102 30 . . PUNCT njb-49 103 1 finally finally ADV njb-49 103 2 , , PUNCT njb-49 103 3 cloned cloned ADJ njb-49 103 4 cell cell NOUN njb-49 103 5 lines line NOUN njb-49 103 6 were be AUX njb-49 103 7 grown grow VERB njb-49 103 8 in in ADP njb-49 103 9 dmem dmem PROPN njb-49 103 10 supplemented supplement VERB njb-49 103 11 with with ADP njb-49 103 12 10 10 NUM njb-49 103 13 % % NOUN njb-49 103 14 fbs fb NOUN njb-49 103 15 and and CCONJ njb-49 103 16 stored store VERB njb-49 103 17 in in ADP njb-49 103 18 liquid liquid ADJ njb-49 103 19 nitrogen nitrogen PROPN njb-49 103 20 . . PUNCT njb-49 104 1 indirect indirect ADJ njb-49 104 2 enzyme enzyme NOUN njb-49 104 3 linked link VERB njb-49 104 4 immunosorbent immunosorbent NOUN njb-49 104 5 assay assay NOUN njb-49 104 6 ( ( PUNCT njb-49 104 7 elisa elisa NOUN njb-49 104 8 ) ) PUNCT njb-49 104 9 for for ADP njb-49 104 10 the the DET njb-49 104 11 measurement measurement NOUN njb-49 104 12 of of ADP njb-49 104 13 serum serum NOUN njb-49 104 14 antibody antibody NOUN njb-49 104 15 titers titer NOUN njb-49 104 16 , , PUNCT njb-49 104 17 elisa elisa PROPN njb-49 104 18 was be AUX njb-49 104 19 performed perform VERB njb-49 104 20 as as ADP njb-49 104 21 a a DET njb-49 104 22 standard standard ADJ njb-49 104 23 protocol protocol NOUN njb-49 104 24 mentioned mention VERB njb-49 104 25 by by ADP njb-49 104 26 sigma sigma PROPN njb-49 104 27 aldrich aldrich PROPN njb-49 104 28 , , PUNCT njb-49 104 29 with with ADP njb-49 104 30 only only ADJ njb-49 104 31 minor minor ADJ njb-49 104 32 modifications modification NOUN njb-49 104 33 . . PUNCT njb-49 105 1 each each DET njb-49 105 2 well well NOUN njb-49 105 3 of of ADP njb-49 105 4 96 96 NUM njb-49 105 5 well well ADJ njb-49 105 6 plates plate NOUN njb-49 105 7 ( ( PUNCT njb-49 105 8 nunc nunc PROPN njb-49 105 9 , , PUNCT njb-49 105 10 copenhagen copenhagen PROPN njb-49 105 11 , , PUNCT njb-49 105 12 denmark denmark PROPN njb-49 105 13 ) ) PUNCT njb-49 105 14 was be AUX njb-49 105 15 coated coat VERB njb-49 105 16 with with ADP njb-49 105 17 antigen antigen NOUN njb-49 105 18 ( ( PUNCT njb-49 105 19 100 100 NUM njb-49 105 20 µl µl NOUN njb-49 105 21 containing contain VERB njb-49 105 22 100 100 NUM njb-49 105 23 ng ng NOUN njb-49 105 24 of of ADP njb-49 105 25 either either CCONJ njb-49 105 26 gst gst NOUN njb-49 105 27 free free ADJ njb-49 105 28 recombinant recombinant ADJ njb-49 105 29 protein protein NOUN njb-49 105 30 or or CCONJ njb-49 105 31 100 100 NUM njb-49 105 32 ng ng PROPN njb-49 105 33 bont bont PROPN njb-49 105 34 / / SYM njb-49 105 35 f f PROPN njb-49 105 36 ) ) PUNCT njb-49 105 37 and and CCONJ njb-49 105 38 incubated incubate VERB njb-49 105 39 at at ADP njb-49 105 40 4 4 NUM njb-49 105 41 ° ° NOUN njb-49 105 42 c c NOUN njb-49 105 43 overnight overnight ADV njb-49 105 44 . . PUNCT njb-49 106 1 each each DET njb-49 106 2 well well ADV njb-49 106 3 was be AUX njb-49 106 4 washed wash VERB njb-49 106 5 once once ADV njb-49 106 6 with with ADP njb-49 106 7 200 200 NUM njb-49 106 8 µl µl NOUN njb-49 106 9 of of ADP njb-49 106 10 pbs pbs PROPN njb-49 106 11 containing contain VERB njb-49 106 12 0.05 0.05 NUM njb-49 106 13 % % NOUN njb-49 106 14 tween tween ADP njb-49 106 15 20 20 NUM njb-49 106 16 ( ( PUNCT njb-49 106 17 pbst pbst VERB njb-49 106 18 ) ) PUNCT njb-49 106 19 and and CCONJ njb-49 106 20 blocking block VERB njb-49 106 21 solution solution NOUN njb-49 106 22 ( ( PUNCT njb-49 106 23 100 100 NUM njb-49 106 24 µl µl NOUN njb-49 106 25 1 1 NUM njb-49 106 26 % % NOUN njb-49 106 27 skim skim NOUN njb-49 106 28 milk milk NOUN njb-49 106 29 ) ) PUNCT njb-49 106 30 was be AUX njb-49 106 31 added add VERB njb-49 106 32 and and CCONJ njb-49 106 33 incubated incubate VERB njb-49 106 34 for for ADP njb-49 106 35 30 30 NUM njb-49 106 36 min min NOUN njb-49 106 37 at at ADP njb-49 106 38 37 37 NUM njb-49 106 39 ° ° NOUN njb-49 106 40 c c NOUN njb-49 106 41 . . PUNCT njb-49 107 1 following follow VERB njb-49 107 2 two two NUM njb-49 107 3 more more ADJ njb-49 107 4 washes wash NOUN njb-49 107 5 , , PUNCT njb-49 107 6 the the DET njb-49 107 7 serum serum NOUN njb-49 107 8 ( ( PUNCT njb-49 107 9 1 1 NUM njb-49 107 10 : : SYM njb-49 107 11 1000 1000 NUM njb-49 107 12 dilution dilution NOUN njb-49 107 13 ) ) PUNCT njb-49 107 14 in in ADP njb-49 107 15 pbs pbs PROPN njb-49 107 16 was be AUX njb-49 107 17 added add VERB njb-49 107 18 as as ADP njb-49 107 19 the the DET njb-49 107 20 primary primary ADJ njb-49 107 21 antibody antibody NOUN njb-49 107 22 . . PUNCT njb-49 108 1 the the DET njb-49 108 2 plates plate NOUN njb-49 108 3 were be AUX njb-49 108 4 incubated incubate VERB njb-49 108 5 at at ADP njb-49 108 6 37 37 NUM njb-49 108 7 ° ° NOUN njb-49 108 8 c c NOUN njb-49 108 9 for for ADP njb-49 108 10 90 90 NUM njb-49 108 11 min min NOUN njb-49 108 12 , , PUNCT njb-49 108 13 and and CCONJ njb-49 108 14 then then ADV njb-49 108 15 washed wash VERB njb-49 108 16 three three NUM njb-49 108 17 times time NOUN njb-49 108 18 as as SCONJ njb-49 108 19 described describe VERB njb-49 108 20 above above ADP njb-49 108 21 . . PUNCT njb-49 109 1 goat goat PROPN njb-49 109 2 anti anti ADJ njb-49 109 3 - - ADJ njb-49 109 4 mouse mouse ADJ njb-49 109 5 igg igg NOUN njb-49 109 6 conjugated conjugate VERB njb-49 109 7 with with ADP njb-49 109 8 alkaline alkaline NOUN njb-49 109 9 phosphatase phosphatase NOUN njb-49 109 10 was be AUX njb-49 109 11 added add VERB njb-49 109 12 as as ADP njb-49 109 13 the the DET njb-49 109 14 secondary secondary ADJ njb-49 109 15 antibody antibody NOUN njb-49 109 16 ( ( PUNCT njb-49 109 17 diluted dilute VERB njb-49 109 18 1:2000 1:2000 NUM njb-49 109 19 with with ADP njb-49 109 20 pbs pbs PROPN njb-49 109 21 ) ) PUNCT njb-49 109 22 and and CCONJ njb-49 109 23 incubated incubate VERB njb-49 109 24 for for ADP njb-49 109 25 2 2 NUM njb-49 109 26 h. h. NOUN njb-49 109 27 after after ADP njb-49 109 28 incubation incubation NOUN njb-49 109 29 with with ADP njb-49 109 30 igg igg PROPN njb-49 109 31 , , PUNCT njb-49 109 32 plates plate NOUN njb-49 109 33 were be AUX njb-49 109 34 washed wash VERB njb-49 109 35 four four NUM njb-49 109 36 times time NOUN njb-49 109 37 prior prior ADV njb-49 109 38 to to ADP njb-49 109 39 substrate substrate ADJ njb-49 109 40 treatment treatment NOUN njb-49 109 41 and and CCONJ njb-49 109 42 visualization visualization NOUN njb-49 109 43 . . PUNCT njb-49 110 1 immobilized immobilized ADJ njb-49 110 2 antigens antigen NOUN njb-49 110 3 were be AUX njb-49 110 4 visualized visualize VERB njb-49 110 5 with with ADP njb-49 110 6 the the DET njb-49 110 7 p p PROPN njb-49 110 8 - - PUNCT njb-49 110 9 nitrophenyl nitrophenyl NOUN njb-49 110 10 phosphate phosphate NOUN njb-49 110 11 ( ( PUNCT njb-49 110 12 disodium disodium NOUN njb-49 110 13 ) ) PUNCT njb-49 110 14 in in ADP njb-49 110 15 substrate substrate ADJ njb-49 110 16 buffer buffer NOUN njb-49 110 17 containing contain VERB njb-49 110 18 9.7 9.7 NUM njb-49 110 19 % % NOUN njb-49 110 20 diethanolamine diethanolamine NOUN njb-49 110 21 , , PUNCT njb-49 110 22 and and CCONJ njb-49 110 23 1 1 NUM njb-49 110 24 mm mm NOUN njb-49 110 25 mgcl2 mgcl2 NOUN njb-49 110 26 and and CCONJ njb-49 110 27 the the DET njb-49 110 28 resultant resultant NOUN njb-49 110 29 absorbance absorbance NOUN njb-49 110 30 was be AUX njb-49 110 31 measured measure VERB njb-49 110 32 at at ADP njb-49 110 33 415 415 NUM njb-49 110 34 nm nm NOUN njb-49 110 35 with with ADP njb-49 110 36 microplate microplate NOUN njb-49 110 37 reader reader NOUN njb-49 110 38 ( ( PUNCT njb-49 110 39 model model NOUN njb-49 110 40 550 550 NUM njb-49 110 41 bio bio NOUN njb-49 110 42 - - ADJ njb-49 110 43 rad rad ADJ njb-49 110 44 ) ) PUNCT njb-49 110 45 for for ADP njb-49 110 46 5 5 NUM njb-49 110 47 10 10 NUM njb-49 110 48 min min NOUN njb-49 110 49 at at ADP njb-49 110 50 37 37 NUM njb-49 110 51 ° ° NUM njb-49 110 52 c c NOUN njb-49 110 53 . . PUNCT njb-49 110 54 western western ADJ njb-49 110 55 blot blot NOUN njb-49 110 56 analysis analysis NOUN njb-49 110 57 western western ADJ njb-49 110 58 blot blot NOUN njb-49 110 59 experimentation experimentation NOUN njb-49 110 60 was be AUX njb-49 110 61 adapted adapt VERB njb-49 110 62 from from ADP njb-49 110 63 previous previous ADJ njb-49 110 64 methodology methodology NOUN njb-49 110 65 [ [ PUNCT njb-49 110 66 26 26 NUM njb-49 110 67 ] ] PUNCT njb-49 110 68 . . PUNCT njb-49 111 1 the the DET njb-49 111 2 protein protein NOUN njb-49 111 3 samples sample NOUN njb-49 111 4 were be AUX njb-49 111 5 separated separate VERB njb-49 111 6 by by ADP njb-49 111 7 sds sds NOUN njb-49 111 8 - - PUNCT njb-49 111 9 page page NOUN njb-49 111 10 and and CCONJ njb-49 111 11 electroblotted electroblotte VERB njb-49 111 12 for for ADP njb-49 111 13 ~70 ~70 NUM njb-49 111 14 min min NOUN njb-49 111 15 by by ADP njb-49 111 16 using use VERB njb-49 111 17 230 230 NUM njb-49 111 18 ma ma PROPN njb-49 111 19 current current NOUN njb-49 111 20 onto onto ADP njb-49 111 21 a a DET njb-49 111 22 nitrocellulose nitrocellulose NOUN njb-49 111 23 membrane membrane NOUN njb-49 111 24 ( ( PUNCT njb-49 111 25 schleicher schleicher NOUN njb-49 111 26 and and CCONJ njb-49 111 27 schuell schuell PROPN njb-49 111 28 inc inc PROPN njb-49 111 29 . . PROPN njb-49 111 30 , , PUNCT njb-49 111 31 nh nh PROPN njb-49 111 32 , , PUNCT njb-49 111 33 usa usa PROPN njb-49 111 34 ) ) PUNCT njb-49 111 35 . . PUNCT njb-49 112 1 nepal nepal ADJ njb-49 112 2     SPACE njb-49 112 3 journal journal NOUN njb-49 112 4     SPACE njb-49 112 5 of of ADP njb-49 112 6     SPACE njb-49 112 7 biotechnology biotechnology NOUN njb-49 112 8 . . PUNCT njb-49 112 9         SPACE njb-49 113 1 jan jan PROPN njb-49 113 2 . . PROPN njb-49 113 3     SPACE njb-49 113 4 2012 2012 NUM njb-49 113 5 , , PUNCT njb-49 113 6     SPACE njb-49 113 7 vol vol NOUN njb-49 113 8 . . PUNCT njb-49 113 9     SPACE njb-49 113 10 2 2 NUM njb-49 113 11 , , PUNCT njb-49 113 12     SPACE njb-49 113 13 no no INTJ njb-49 113 14 . . PUNCT njb-49 113 15     SPACE njb-49 114 1 1 1 NUM njb-49 114 2 : : PUNCT njb-49 114 3     SPACE njb-49 114 4 1 1 X njb-49 114 5     SPACE njb-49 114 6 – – PUNCT njb-49 114 7     SPACE njb-49 114 8 15 15 NUM njb-49 114 9                                                                                                                                                                                                                                         SPACE njb-49 114 10 biotechnology biotechnology NOUN njb-49 114 11     SPACE njb-49 114 12 society society NOUN njb-49 114 13     SPACE njb-49 114 14 of of ADP njb-49 114 15     SPACE njb-49 114 16 nepal nepal NOUN njb-49 114 17     SPACE njb-49 114 18 ( ( PUNCT njb-49 114 19 bsn bsn PROPN njb-49 114 20 ) ) PUNCT njb-49 114 21 , , PUNCT njb-49 114 22     SPACE njb-49 114 23 all all DET njb-49 114 24     SPACE njb-49 114 25 rights right NOUN njb-49 114 26     SPACE njb-49 114 27 reserved reserve VERB njb-49 114 28     SPACE njb-49 114 29 7 7 NUM njb-49 114 30 subsequently subsequently ADV njb-49 114 31 , , PUNCT njb-49 114 32 the the DET njb-49 114 33 membrane membrane NOUN njb-49 114 34 was be AUX njb-49 114 35 blocked block VERB njb-49 114 36 for for ADP njb-49 114 37 nonspecific nonspecific ADJ njb-49 114 38 binding bind VERB njb-49 114 39 incubating incubate VERB njb-49 114 40 with with ADP njb-49 114 41 blocking block VERB njb-49 114 42 solution solution NOUN njb-49 114 43 for for ADP njb-49 114 44 1 1 NUM njb-49 114 45 h h NOUN njb-49 114 46 and and CCONJ njb-49 114 47 then then ADV njb-49 114 48 incubated incubate VERB njb-49 114 49 with with ADP njb-49 114 50 primary primary ADJ njb-49 114 51 antibody antibody NOUN njb-49 114 52 overnight overnight ADV njb-49 114 53 at at ADP njb-49 114 54 room room NOUN njb-49 114 55 temp temp NOUN njb-49 114 56 . . PUNCT njb-49 115 1 the the DET njb-49 115 2 membrane membrane NOUN njb-49 115 3 was be AUX njb-49 115 4 washed wash VERB njb-49 115 5 two two NUM njb-49 115 6 times time NOUN njb-49 115 7 with with ADP njb-49 115 8 pbst pbst NOUN njb-49 115 9 and and CCONJ njb-49 115 10 incubated incubate VERB njb-49 115 11 with with ADP njb-49 115 12 alkaline alkaline NOUN njb-49 115 13 phosphate phosphate NOUN njb-49 115 14 conjugated conjugate VERB njb-49 115 15 goat goat PROPN njb-49 115 16 anti anti ADJ njb-49 115 17 - - ADJ njb-49 115 18 mouse mouse ADJ njb-49 115 19 igg igg PROPN njb-49 115 20 as as ADP njb-49 115 21 a a DET njb-49 115 22 secondary secondary ADJ njb-49 115 23 antibody antibody NOUN njb-49 115 24 for for ADP njb-49 115 25 2 2 NUM njb-49 115 26 h. h. NOUN njb-49 115 27 after after ADP njb-49 115 28 washing wash VERB njb-49 115 29 three three NUM njb-49 115 30 more more ADJ njb-49 115 31 times time NOUN njb-49 115 32 with with ADP njb-49 115 33 pbst pbst NOUN njb-49 115 34 , , PUNCT njb-49 115 35 the the DET njb-49 115 36 membrane membrane NOUN njb-49 115 37 was be AUX njb-49 115 38 color color NOUN njb-49 115 39 developed develop VERB njb-49 115 40 by by ADP njb-49 115 41 using use VERB njb-49 115 42 10 10 NUM njb-49 115 43 ml ml PROPN njb-49 115 44 alkaline alkaline NOUN njb-49 115 45 phosphate phosphate NOUN njb-49 115 46 substrate substrate NOUN njb-49 115 47 solution solution NOUN njb-49 115 48 , , PUNCT njb-49 115 49 50 50 NUM njb-49 115 50 µl µl PROPN njb-49 115 51 nitro nitro PROPN njb-49 115 52 blue blue PROPN njb-49 115 53 tetrazolium tetrazolium PROPN njb-49 115 54 chloride chloride PROPN njb-49 115 55 , , PUNCT njb-49 115 56 and and CCONJ njb-49 115 57 50 50 NUM njb-49 115 58 µl µl NOUN njb-49 115 59 5 5 NUM njb-49 115 60 - - PUNCT njb-49 115 61 bromo-4chloro-3 bromo-4chloro-3 ADJ njb-49 115 62 - - PUNCT njb-49 115 63 indolyl indolyl PROPN njb-49 115 64 phosphate phosphate NOUN njb-49 115 65 . . PUNCT njb-49 116 1 results result NOUN njb-49 116 2 and and CCONJ njb-49 116 3 discussion discussion VERB njb-49 116 4 the the DET njb-49 116 5 cloning cloning NOUN njb-49 116 6 of of ADP njb-49 116 7 both both PRON njb-49 116 8 light light ADJ njb-49 116 9 and and CCONJ njb-49 116 10 heavy heavy ADJ njb-49 116 11 chain chain NOUN njb-49 116 12 domains domain NOUN njb-49 116 13 using use VERB njb-49 116 14 synthetic synthetic ADJ njb-49 116 15 genes gene NOUN njb-49 116 16 has have AUX njb-49 116 17 been be AUX njb-49 116 18 reported report VERB njb-49 116 19 for for ADP njb-49 116 20 various various ADJ njb-49 116 21 bonts bont NOUN njb-49 116 22 [ [ PUNCT njb-49 116 23 27 27 NUM njb-49 116 24 - - SYM njb-49 116 25 33 33 NUM njb-49 116 26 ] ] PUNCT njb-49 116 27 . . PUNCT njb-49 117 1 however however ADV njb-49 117 2 , , PUNCT njb-49 117 3 there there PRON njb-49 117 4 exist exist VERB njb-49 117 5 few few ADJ njb-49 117 6 studies study NOUN njb-49 117 7 on on ADP njb-49 117 8 the the DET njb-49 117 9 cloning cloning NOUN njb-49 117 10 and and CCONJ njb-49 117 11 expression expression NOUN njb-49 117 12 of of ADP njb-49 117 13 bont bont PROPN njb-49 117 14 / / SYM njb-49 117 15 f f NOUN njb-49 117 16 domains domain NOUN njb-49 117 17 [ [ PUNCT njb-49 117 18 34 34 NUM njb-49 117 19 , , PUNCT njb-49 117 20 35 35 NUM njb-49 117 21 ] ] PUNCT njb-49 117 22 . . PUNCT njb-49 118 1 construction construction NOUN njb-49 118 2 of of ADP njb-49 118 3 recombinant recombinant ADJ njb-49 118 4 plasmids plasmid NOUN njb-49 118 5 containing contain VERB njb-49 118 6 pgex-4t-1 pgex-4t-1 PROPN njb-49 118 7 - - PUNCT njb-49 118 8 f f PROPN njb-49 118 9 - - PUNCT njb-49 118 10 lc lc NOUN njb-49 118 11 , , PUNCT njb-49 118 12 -hn -hn PUNCT njb-49 118 13 , , PUNCT njb-49 118 14 -hc -hc PROPN njb-49 118 15 , , PUNCT njb-49 118 16 hcc hcc PROPN njb-49 118 17 was be AUX njb-49 118 18 made make VERB njb-49 118 19 for for ADP njb-49 118 20 subsequent subsequent ADJ njb-49 118 21 gst gst NOUN njb-49 118 22 - - PUNCT njb-49 118 23 tagged tag VERB njb-49 118 24 expression expression NOUN njb-49 118 25 in in ADP njb-49 118 26 e. e. PROPN njb-49 118 27 coli coli PROPN njb-49 118 28 . . PUNCT njb-49 119 1 initially initially ADV njb-49 119 2 , , PUNCT njb-49 119 3 sequences sequence NOUN njb-49 119 4 of of ADP njb-49 119 5 bont bont PROPN njb-49 119 6 / / SYM njb-49 119 7 f f PROPN njb-49 119 8 dna dna PROPN njb-49 119 9 domains domain NOUN njb-49 119 10 were be AUX njb-49 119 11 pcr pcr NOUN njb-49 119 12 amplified amplify VERB njb-49 119 13 from from ADP njb-49 119 14 genomic genomic ADJ njb-49 119 15 dna dna PROPN njb-49 119 16 of of ADP njb-49 119 17 c. c. PROPN njb-49 119 18 botulinum botulinum PROPN njb-49 119 19 f f PROPN njb-49 119 20 str str PROPN njb-49 119 21 . . PUNCT njb-49 119 22 langeland langeland PROPN njb-49 119 23 and and CCONJ njb-49 119 24 flanked flank VERB njb-49 119 25 by by ADP njb-49 119 26 restriction restriction NOUN njb-49 119 27 sites site NOUN njb-49 119 28 ( ( PUNCT njb-49 119 29 table table NOUN njb-49 119 30 1 1 NUM njb-49 119 31 ) ) PUNCT njb-49 119 32 . . PUNCT njb-49 120 1 specifically specifically ADV njb-49 120 2 , , PUNCT njb-49 120 3 the the DET njb-49 120 4 amplified amplified ADJ njb-49 120 5 products product NOUN njb-49 120 6 were be AUX njb-49 120 7 cloned clone VERB njb-49 120 8 into into ADP njb-49 120 9 pgex-4t-1 pgex-4t-1 PROPN njb-49 120 10 after after ADP njb-49 120 11 digestion digestion NOUN njb-49 120 12 with with ADP njb-49 120 13 respective respective ADJ njb-49 120 14 restriction restriction NOUN njb-49 120 15 enzymes enzyme NOUN njb-49 120 16 . . PUNCT njb-49 121 1 ligated ligate VERB njb-49 121 2 constructs construct NOUN njb-49 121 3 ( ( PUNCT njb-49 121 4 for for ADP njb-49 121 5 reference reference NOUN njb-49 121 6 see see VERB njb-49 121 7 schematic schematic ADJ njb-49 121 8 diagrams diagram NOUN njb-49 121 9 of of ADP njb-49 121 10 constructed construct VERB njb-49 121 11 plasmids plasmid NOUN njb-49 121 12 made make VERB njb-49 121 13 by by ADP njb-49 121 14 macvector macvector NOUN njb-49 121 15 software software NOUN njb-49 121 16 10.0.2 10.0.2 NUM njb-49 121 17 , , PUNCT njb-49 121 18 fig fig NOUN njb-49 121 19 . . PROPN njb-49 121 20 2 2 NUM njb-49 121 21 ) ) PUNCT njb-49 121 22 were be AUX njb-49 121 23 transformed transform VERB njb-49 121 24 into into ADP njb-49 121 25 dh5α dh5α PROPN njb-49 121 26 for for ADP njb-49 121 27 propagation propagation NOUN njb-49 121 28 and and CCONJ njb-49 121 29 positive positive ADJ njb-49 121 30 colonies colony NOUN njb-49 121 31 were be AUX njb-49 121 32 identified identify VERB njb-49 121 33 after after ADP njb-49 121 34 screening screen VERB njb-49 121 35 on on ADP njb-49 121 36 plates plate NOUN njb-49 121 37 containing contain VERB njb-49 121 38 ampicillin ampicillin NOUN njb-49 121 39 . . PUNCT njb-49 122 1 subsequently subsequently ADV njb-49 122 2 the the DET njb-49 122 3 recombinant recombinant ADJ njb-49 122 4 plasmids plasmid NOUN njb-49 122 5 were be AUX njb-49 122 6 digested digest VERB njb-49 122 7 with with ADP njb-49 122 8 restriction restriction NOUN njb-49 122 9 enzymes enzyme NOUN njb-49 122 10 ( ( PUNCT njb-49 122 11 table table NOUN njb-49 122 12 1 1 NUM njb-49 122 13 ) ) PUNCT njb-49 122 14 and and CCONJ njb-49 122 15 appropriate appropriate ADJ njb-49 122 16 band band NOUN njb-49 122 17 sizes size NOUN njb-49 122 18 were be AUX njb-49 122 19 identified identify VERB njb-49 122 20 for for ADP njb-49 122 21 both both PRON njb-49 122 22 pgex-4t-1 pgex-4t-1 ADJ njb-49 122 23 ( ( PUNCT njb-49 122 24 4.9 4.9 NUM njb-49 122 25 kb kb NOUN njb-49 122 26 ) ) PUNCT njb-49 122 27 and and CCONJ njb-49 122 28 all all DET njb-49 122 29 individual individual ADJ njb-49 122 30 fragment fragment ADJ njb-49 122 31 sizes size NOUN njb-49 122 32 ( ( PUNCT njb-49 122 33 fig fig NOUN njb-49 122 34 . . PUNCT njb-49 122 35 3 3 NUM njb-49 122 36 ) ) PUNCT njb-49 122 37 . . PUNCT njb-49 123 1 for for ADP njb-49 123 2 confirmation confirmation NOUN njb-49 123 3 of of ADP njb-49 123 4 domain domain NOUN njb-49 123 5 homology homology NOUN njb-49 123 6 incorporated incorporate VERB njb-49 123 7 into into ADP njb-49 123 8 our our PRON njb-49 123 9 vector vector NOUN njb-49 123 10 , , PUNCT njb-49 123 11 sequencing sequence VERB njb-49 123 12 was be AUX njb-49 123 13 performed perform VERB njb-49 123 14 . . PUNCT njb-49 124 1 after after ADP njb-49 124 2 sequence sequence NOUN njb-49 124 3 analysis analysis NOUN njb-49 124 4 , , PUNCT njb-49 124 5 plasmid plasmid PROPN njb-49 124 6 dna dna NOUN njb-49 124 7 of of ADP njb-49 124 8 positive positive ADJ njb-49 124 9 clones clone NOUN njb-49 124 10 had have VERB njb-49 124 11 100 100 NUM njb-49 124 12 % % NOUN njb-49 124 13 identity identity NOUN njb-49 124 14 with with ADP njb-49 124 15 native native ADJ njb-49 124 16 sequences sequence NOUN njb-49 124 17 ( ( PUNCT njb-49 124 18 data datum NOUN njb-49 124 19 not not PART njb-49 124 20 shown show VERB njb-49 124 21 ) ) PUNCT njb-49 124 22 . . PUNCT njb-49 125 1 for for ADP njb-49 125 2 protein protein NOUN njb-49 125 3 expression expression NOUN njb-49 125 4 , , PUNCT njb-49 125 5 the the DET njb-49 125 6 four four NUM njb-49 125 7 recombinant recombinant ADJ njb-49 125 8 clones clone NOUN njb-49 125 9 were be AUX njb-49 125 10 transformed transform VERB njb-49 125 11 into into ADP njb-49 125 12 e. e. PROPN njb-49 125 13 coli coli PROPN njb-49 125 14 bl21codonplus bl21codonplus NOUN njb-49 125 15 - - PUNCT njb-49 125 16 ril ril NOUN njb-49 125 17 ( ( PUNCT njb-49 125 18 strain strain NOUN njb-49 125 19 used use VERB njb-49 125 20 for for ADP njb-49 125 21 protein protein NOUN njb-49 125 22 expression expression NOUN njb-49 125 23 with with ADP njb-49 125 24 vectors vector NOUN njb-49 125 25 driven drive VERB njb-49 125 26 by by ADP njb-49 125 27 non non ADJ njb-49 125 28 - - ADJ njb-49 125 29 t7 t7 ADJ njb-49 125 30 promoters promoter NOUN njb-49 125 31 ) ) PUNCT njb-49 125 32 . . PUNCT njb-49 126 1 the the DET njb-49 126 2 bl21 bl21 NOUN njb-49 126 3 - - PUNCT njb-49 126 4 codonplus codonplus NOUN njb-49 126 5 - - PUNCT njb-49 126 6 ril ril NOUN njb-49 126 7 was be AUX njb-49 126 8 chosen choose VERB njb-49 126 9 because because SCONJ njb-49 126 10 of of ADP njb-49 126 11 its its PRON njb-49 126 12 capability capability NOUN njb-49 126 13 to to PART njb-49 126 14 express express VERB njb-49 126 15 rare rare ADJ njb-49 126 16 codons codon NOUN njb-49 126 17 which which PRON njb-49 126 18 allows allow VERB njb-49 126 19 for for ADP njb-49 126 20 the the DET njb-49 126 21 high high ADJ njb-49 126 22 - - PUNCT njb-49 126 23 level level NOUN njb-49 126 24 expression expression NOUN njb-49 126 25 of of ADP njb-49 126 26 recombinants recombinant NOUN njb-49 126 27 [ [ PUNCT njb-49 126 28 36 36 NUM njb-49 126 29 ] ] PUNCT njb-49 126 30 . . PUNCT njb-49 127 1 thus thus ADV njb-49 127 2 , , PUNCT njb-49 127 3 bl21codonplus bl21codonplus NOUN njb-49 127 4 - - PUNCT njb-49 127 5 ril ril NOUN njb-49 127 6 was be AUX njb-49 127 7 a a DET njb-49 127 8 compatible compatible ADJ njb-49 127 9 host host NOUN njb-49 127 10 strain strain NOUN njb-49 127 11 for for ADP njb-49 127 12 our our PRON njb-49 127 13 pgex-4t-1 pgex-4t-1 ADJ njb-49 127 14 plasmid plasmid NOUN njb-49 127 15 systems system NOUN njb-49 127 16 ( ( PUNCT njb-49 127 17 containing contain VERB njb-49 127 18 a a DET njb-49 127 19 tac tac NOUN njb-49 127 20 promoter promoter NOUN njb-49 127 21 ) ) PUNCT njb-49 127 22 . . PUNCT njb-49 128 1 all all DET njb-49 128 2 domains domain NOUN njb-49 128 3 were be AUX njb-49 128 4 expressed express VERB njb-49 128 5 in in ADP njb-49 128 6 the the DET njb-49 128 7 form form NOUN njb-49 128 8 of of ADP njb-49 128 9 fusion fusion NOUN njb-49 128 10 proteins protein NOUN njb-49 128 11 with with ADP njb-49 128 12 gst gst NOUN njb-49 128 13 localized localize VERB njb-49 128 14 at at ADP njb-49 128 15 the the DET njb-49 128 16 n n NOUN njb-49 128 17 - - NOUN njb-49 128 18 terminal terminal NOUN njb-49 128 19 of of ADP njb-49 128 20 the the DET njb-49 128 21 fusion fusion NOUN njb-49 128 22 protein protein NOUN njb-49 128 23 , , PUNCT njb-49 128 24 enabling enable VERB njb-49 128 25 the the DET njb-49 128 26 nepal nepal ADJ njb-49 128 27     SPACE njb-49 128 28 journal journal NOUN njb-49 128 29     SPACE njb-49 128 30 of of ADP njb-49 128 31     SPACE njb-49 128 32 biotechnology biotechnology NOUN njb-49 128 33 . . PUNCT njb-49 128 34         SPACE njb-49 129 1 jan jan PROPN njb-49 129 2 . . PROPN njb-49 129 3     SPACE njb-49 129 4 2012 2012 NUM njb-49 129 5 , , PUNCT njb-49 129 6     SPACE njb-49 129 7 vol vol NOUN njb-49 129 8 . . PUNCT njb-49 129 9     SPACE njb-49 129 10 2 2 NUM njb-49 129 11 , , PUNCT njb-49 129 12     SPACE njb-49 129 13 no no INTJ njb-49 129 14 . . PUNCT njb-49 129 15     SPACE njb-49 130 1 1 1 NUM njb-49 130 2 : : PUNCT njb-49 130 3     SPACE njb-49 130 4 1 1 X njb-49 130 5     SPACE njb-49 130 6 – – PUNCT njb-49 130 7     SPACE njb-49 130 8 15 15 NUM njb-49 130 9                                                                                                                                                                                                                                         SPACE njb-49 130 10 biotechnology biotechnology NOUN njb-49 130 11     SPACE njb-49 130 12 society society NOUN njb-49 130 13     SPACE njb-49 130 14 of of ADP njb-49 130 15     SPACE njb-49 130 16 nepal nepal NOUN njb-49 130 17     SPACE njb-49 130 18 ( ( PUNCT njb-49 130 19 bsn bsn PROPN njb-49 130 20 ) ) PUNCT njb-49 130 21 , , PUNCT njb-49 130 22     SPACE njb-49 130 23 all all DET njb-49 130 24     SPACE njb-49 130 25 rights right NOUN njb-49 130 26     SPACE njb-49 130 27 reserved reserve VERB njb-49 130 28     SPACE njb-49 130 29 8 8 NUM njb-49 130 30 ( ( PUNCT njb-49 130 31 i i NOUN njb-49 130 32 ) ) PUNCT njb-49 130 33 visible visible ADJ njb-49 130 34 bands band NOUN njb-49 130 35 of of ADP njb-49 130 36 amplified amplify VERB njb-49 130 37 f f PROPN njb-49 130 38 - - PUNCT njb-49 130 39 lc lc NOUN njb-49 130 40 product product NOUN njb-49 130 41 ( ( PUNCT njb-49 130 42 3a 3a NUM njb-49 130 43 ) ) PUNCT njb-49 130 44 , , PUNCT njb-49 130 45 f f X njb-49 130 46 - - PUNCT njb-49 130 47 hn hn PROPN njb-49 130 48 ( ( PUNCT njb-49 130 49 3b 3b NUM njb-49 130 50 ) ) PUNCT njb-49 130 51 , , PUNCT njb-49 130 52 f f X njb-49 130 53 - - PUNCT njb-49 130 54 hc hc PROPN njb-49 130 55 ( ( PUNCT njb-49 130 56 3c 3c NUM njb-49 130 57 ) ) PUNCT njb-49 130 58 , , PUNCT njb-49 130 59 and and CCONJ njb-49 130 60 f f X njb-49 130 61 - - NOUN njb-49 130 62 hcc hcc PROPN njb-49 130 63 ( ( PUNCT njb-49 130 64 3d 3d NOUN njb-49 130 65 ) ) PUNCT njb-49 130 66 ; ; PUNCT njb-49 130 67 ( ( PUNCT njb-49 130 68 ii ii X njb-49 130 69 ) ) PUNCT njb-49 130 70 endonuclease endonuclease NOUN njb-49 130 71 restricted restrict VERB njb-49 130 72 bands band NOUN njb-49 130 73 of of ADP njb-49 130 74 purified purified ADJ njb-49 130 75 pgex4t-1 pgex4t-1 PROPN njb-49 130 76 - - PUNCT njb-49 130 77 f f NOUN njb-49 130 78 - - PUNCT njb-49 130 79 lc lc NOUN njb-49 130 80 with with ADP njb-49 130 81 sali sali PROPN njb-49 130 82 / / SYM njb-49 130 83 noti noti PROPN njb-49 130 84 ( ( PUNCT njb-49 130 85 3a 3a NUM njb-49 130 86 ) ) PUNCT njb-49 130 87 , , PUNCT njb-49 130 88 pgex-4t-1 pgex-4t-1 PROPN njb-49 130 89 - - PUNCT njb-49 130 90 f f PROPN njb-49 130 91 - - PUNCT njb-49 130 92 hn hn NOUN njb-49 130 93 ( ( PUNCT njb-49 130 94 3b 3b NUM njb-49 130 95 ) ) PUNCT njb-49 130 96 , , PUNCT njb-49 130 97 pgex-4t-1 pgex-4t-1 PROPN njb-49 130 98 - - PUNCT njb-49 130 99 f f PROPN njb-49 130 100 - - PUNCT njb-49 130 101 hc hc PROPN njb-49 130 102 ( ( PUNCT njb-49 130 103 3c 3c NUM njb-49 130 104 ) ) PUNCT njb-49 130 105 , , PUNCT njb-49 130 106 and and CCONJ njb-49 130 107 pgex-4t-1 pgex-4t-1 PROPN njb-49 130 108 - - PUNCT njb-49 130 109 f f PROPN njb-49 130 110 - - PUNCT njb-49 130 111 hcc hcc PROPN njb-49 130 112 ( ( PUNCT njb-49 130 113 3d 3d NOUN njb-49 130 114 ) ) PUNCT njb-49 130 115 with with ADP njb-49 130 116 bami bami PROPN njb-49 130 117 / / SYM njb-49 130 118 xhoi xhoi PROPN njb-49 130 119 . . PUNCT njb-49 131 1 m m VERB njb-49 131 2 denotes denote VERB njb-49 131 3 dna dna PROPN njb-49 131 4 marker marker NOUN njb-49 131 5 and and CCONJ njb-49 131 6 lane lane NOUN njb-49 131 7 1 1 NUM njb-49 131 8 , , PUNCT njb-49 131 9 dna dna PROPN njb-49 131 10 sample sample NOUN njb-49 131 11 loaded load VERB njb-49 131 12 . . PUNCT njb-49 132 1 ease ease NOUN njb-49 132 2 of of ADP njb-49 132 3 purification purification NOUN njb-49 132 4 using use VERB njb-49 132 5 gst gst PROPN njb-49 132 6 affinity affinity NOUN njb-49 132 7 chromatography chromatography NOUN njb-49 132 8 . . PUNCT njb-49 133 1 expression expression NOUN njb-49 133 2 of of ADP njb-49 133 3 clostridial clostridial ADJ njb-49 133 4 proteins protein NOUN njb-49 133 5 at at ADP njb-49 133 6 37 37 NUM njb-49 133 7 ° ° NOUN njb-49 133 8 c c PROPN njb-49 133 9 has have AUX njb-49 133 10 been be AUX njb-49 133 11 shown show VERB njb-49 133 12 to to PART njb-49 133 13 increase increase VERB njb-49 133 14 protein protein NOUN njb-49 133 15 degradation degradation NOUN njb-49 133 16 [ [ PUNCT njb-49 133 17 37 37 NUM njb-49 133 18 ] ] PUNCT njb-49 133 19 , , PUNCT njb-49 133 20 thus thus ADV njb-49 133 21 we we PRON njb-49 133 22 chose choose VERB njb-49 133 23 an an DET njb-49 133 24 optimal optimal ADJ njb-49 133 25 temperature temperature NOUN njb-49 133 26 range range NOUN njb-49 133 27 of of ADP njb-49 133 28 20 20 NUM njb-49 133 29 25 25 NUM njb-49 133 30 ° ° NOUN njb-49 133 31 c c NOUN njb-49 133 32 for for ADP njb-49 133 33 our our PRON njb-49 133 34 expression expression NOUN njb-49 133 35 studies study NOUN njb-49 133 36 . . PUNCT njb-49 134 1 conditions condition NOUN njb-49 134 2 for for ADP njb-49 134 3 expression expression NOUN njb-49 134 4 of of ADP njb-49 134 5 recombinant recombinant ADJ njb-49 134 6 proteins protein NOUN njb-49 134 7 were be AUX njb-49 134 8 as as SCONJ njb-49 134 9 follows follow VERB njb-49 134 10 : : PUNCT njb-49 134 11 7 7 NUM njb-49 134 12 h h NOUN njb-49 134 13 at at ADP njb-49 134 14 24 24 NUM njb-49 134 15 ° ° NOUN njb-49 134 16 c c NOUN njb-49 134 17 using use VERB njb-49 134 18 0.5 0.5 NUM njb-49 134 19 mm mm NOUN njb-49 134 20 iptg iptg NOUN njb-49 134 21 concentration concentration NOUN njb-49 134 22 for for ADP njb-49 134 23 gst gst NOUN njb-49 134 24 - - PUNCT njb-49 134 25 f f PROPN njb-49 134 26 - - PUNCT njb-49 134 27 lc lc PROPN njb-49 134 28 , , PUNCT njb-49 134 29 8 8 NUM njb-49 134 30 h h NOUN njb-49 134 31 at at ADP njb-49 134 32 22 22 NUM njb-49 134 33 ° ° NOUN njb-49 134 34 c c NOUN njb-49 134 35 using use VERB njb-49 134 36 0.4 0.4 NUM njb-49 134 37 mm mm NOUN njb-49 134 38 iptg iptg NOUN njb-49 134 39 concentration concentration NOUN njb-49 134 40 for for ADP njb-49 134 41 gst gst NOUN njb-49 134 42 - - PUNCT njb-49 134 43 f f PROPN njb-49 134 44 - - PUNCT njb-49 134 45 hn hn PROPN njb-49 134 46 , , PUNCT njb-49 134 47 8 8 NUM njb-49 134 48 h h NOUN njb-49 134 49 at at ADP njb-49 134 50 20 20 NUM njb-49 134 51 ° ° NOUN njb-49 134 52 c c NOUN njb-49 134 53 using use VERB njb-49 134 54 0.25 0.25 NUM njb-49 134 55 mm mm NOUN njb-49 134 56 iptg iptg NOUN njb-49 134 57 concentration concentration NOUN njb-49 134 58 for for ADP njb-49 134 59 gst gst NOUN njb-49 134 60 - - PUNCT njb-49 134 61 fhc fhc PROPN njb-49 134 62 , , PUNCT njb-49 134 63 and and CCONJ njb-49 134 64 8 8 NUM njb-49 134 65 h h NOUN njb-49 134 66 at at ADP njb-49 134 67 20 20 NUM njb-49 134 68 ° ° NOUN njb-49 134 69 c c NOUN njb-49 134 70 using use VERB njb-49 134 71 0.10 0.10 NUM njb-49 134 72 mm mm NOUN njb-49 134 73 iptg iptg NOUN njb-49 134 74 concentration concentration NOUN njb-49 134 75 for for ADP njb-49 134 76 gst gst NOUN njb-49 134 77 - - PUNCT njb-49 134 78 f f PROPN njb-49 134 79 - - PUNCT njb-49 134 80 hcc hcc PROPN njb-49 134 81 . . PUNCT njb-49 135 1 gst gst PROPN njb-49 135 2 - - PUNCT njb-49 135 3 f f PROPN njb-49 135 4 - - PUNCT njb-49 135 5 lc lc NOUN njb-49 135 6 and and CCONJ njb-49 135 7 gst gst PROPN njb-49 135 8 - - PUNCT njb-49 135 9 f f PROPN njb-49 135 10 - - PUNCT njb-49 135 11 hn hn PROPN njb-49 135 12 induction induction NOUN njb-49 135 13 was be AUX njb-49 135 14 monitored monitor VERB njb-49 135 15 by by ADP njb-49 135 16 sdspage sdspage PROPN njb-49 135 17 ( ( PUNCT njb-49 135 18 fig fig NOUN njb-49 135 19 . . PUNCT njb-49 135 20 4 4 NUM njb-49 135 21 ) ) PUNCT njb-49 135 22 and and CCONJ njb-49 135 23 were be AUX njb-49 135 24 found find VERB njb-49 135 25 to to PART njb-49 135 26 be be AUX njb-49 135 27 highly highly ADV njb-49 135 28 over over ADV njb-49 135 29 - - PUNCT njb-49 135 30 expressed express VERB njb-49 135 31 in in ADP njb-49 135 32 soluble soluble ADJ njb-49 135 33 form form NOUN njb-49 135 34 and and CCONJ njb-49 135 35 purified purify VERB njb-49 135 36 at at ADP njb-49 135 37 high high ADJ njb-49 135 38 concentrations concentration NOUN njb-49 135 39 . . PUNCT njb-49 136 1 despite despite SCONJ njb-49 136 2 induction induction NOUN njb-49 136 3 at at ADP njb-49 136 4 low low ADJ njb-49 136 5 temperature temperature NOUN njb-49 136 6 and and CCONJ njb-49 136 7 iptg iptg NOUN njb-49 136 8 concentrations concentration NOUN njb-49 136 9 , , PUNCT njb-49 136 10 gst gst PROPN njb-49 136 11 - - PUNCT njb-49 136 12 f f PROPN njb-49 136 13 - - PUNCT njb-49 136 14 hc hc PROPN njb-49 136 15 and and CCONJ njb-49 136 16 gst gst PROPN njb-49 136 17 - - PUNCT njb-49 136 18 f f PROPN njb-49 136 19 - - PROPN njb-49 136 20 hcc hcc PROPN njb-49 136 21 were be AUX njb-49 136 22 expressed express VERB njb-49 136 23 in in ADP njb-49 136 24 inclusion inclusion NOUN njb-49 136 25 bodies body NOUN njb-49 136 26 . . PUNCT njb-49 137 1 change change NOUN njb-49 137 2 in in ADP njb-49 137 3 expression expression NOUN njb-49 137 4 vectors vector NOUN njb-49 137 5 from from ADP njb-49 137 6 pgex-4t-1 pgex-4t-1 PROPN njb-49 137 7 to to ADP njb-49 137 8 pet32a(+ pet32a(+ NOUN njb-49 137 9 ) ) PUNCT njb-49 137 10 was be AUX njb-49 137 11 then then ADV njb-49 137 12 carried carry VERB njb-49 137 13 out out ADP njb-49 137 14 in in ADP njb-49 137 15 attempt attempt NOUN njb-49 137 16 to to PART njb-49 137 17 obtain obtain VERB njb-49 137 18 soluble soluble ADJ njb-49 137 19 forms form NOUN njb-49 137 20 of of ADP njb-49 137 21 gst gst NOUN njb-49 137 22 - - PUNCT njb-49 137 23 f f PROPN njb-49 137 24 - - PUNCT njb-49 137 25 hc hc PROPN njb-49 137 26 and and CCONJ njb-49 137 27 gst gst PROPN njb-49 137 28 - - PUNCT njb-49 137 29 f f PROPN njb-49 137 30 - - PUNCT njb-49 137 31 hcc hcc PROPN njb-49 137 32 . . PUNCT njb-49 138 1 pet-32a(+ pet-32a(+ PROPN njb-49 138 2 ) ) PUNCT njb-49 138 3 was be AUX njb-49 138 4 subsequently subsequently ADV njb-49 138 5 transformed transform VERB njb-49 138 6 into into ADP njb-49 138 7 bl21 bl21 NOUN njb-49 138 8 - - PUNCT njb-49 138 9 codonplus(de3)-ril codonplus(de3)-ril NOUN njb-49 138 10 host host NOUN njb-49 138 11 cells cell NOUN njb-49 138 12 , , PUNCT njb-49 138 13 that that PRON njb-49 138 14 utilizes utilize VERB njb-49 138 15 the the DET njb-49 138 16 t7 t7 PROPN njb-49 138 17 rna rna PROPN njb-49 138 18 polymerase polymerase NOUN njb-49 138 19 promoter promoter NOUN njb-49 138 20 . . PUNCT njb-49 139 1 m m VERB njb-49 139 2 1 1 NUM njb-49 139 3 m m SYM njb-49 139 4 1 1 NUM njb-49 139 5 insert insert PROPN njb-49 139 6 f f PROPN njb-49 139 7 - - PUNCT njb-49 139 8 lc lc PROPN njb-49 139 9 ( ( PUNCT njb-49 139 10 1308 1308 NUM njb-49 139 11 bp bp PROPN njb-49 139 12 ) ) PUNCT njb-49 139 13 vector vector NOUN njb-49 139 14 pgex-4t-1 pgex-4t-1 PROPN njb-49 139 15 ( ( PUNCT njb-49 139 16 4.9 4.9 NUM njb-49 139 17 kb kb PROPN njb-49 139 18 ) ) PUNCT njb-49 139 19 bp bp PROPN njb-49 139 20 300 300 NUM njb-49 139 21 400 400 NUM njb-49 139 22 850 850 NUM njb-49 139 23 650 650 NUM njb-49 139 24 1650 1650 NUM njb-49 139 25 3000 3000 NUM njb-49 139 26 2000 2000 NUM njb-49 139 27 4000 4000 NUM njb-49 139 28 5000 5000 NUM njb-49 139 29 500 500 NUM njb-49 139 30 1000 1000 NUM njb-49 139 31 100 100 NUM njb-49 139 32 400 400 NUM njb-49 139 33 500 500 NUM njb-49 139 34 3000 3000 NUM njb-49 139 35 2000 2000 NUM njb-49 139 36 5000 5000 NUM njb-49 139 37 1000 1000 NUM njb-49 139 38 300 300 NUM njb-49 139 39 200 200 NUM njb-49 139 40 650 650 NUM njb-49 139 41 850 850 NUM njb-49 139 42 1650 1650 NUM njb-49 139 43 4000 4000 NUM njb-49 139 44 amplified amplify VERB njb-49 139 45 f f PROPN njb-49 139 46 - - PUNCT njb-49 139 47 lc lc NOUN njb-49 139 48 pcr pcr NOUN njb-49 139 49 product product NOUN njb-49 139 50 ( ( PUNCT njb-49 139 51 1308 1308 NUM njb-49 139 52 bp bp PROPN njb-49 139 53 ) ) PUNCT njb-49 139 54 a a PRON njb-49 139 55 ( ( PUNCT njb-49 139 56 i i NOUN njb-49 139 57 ) ) PUNCT njb-49 139 58 bp bp PROPN njb-49 139 59 a a DET njb-49 139 60 ( ( PUNCT njb-49 139 61 ii ii PROPN njb-49 139 62 ) ) PUNCT njb-49 139 63 amplified amplify VERB njb-49 139 64 f f PROPN njb-49 139 65 - - PROPN njb-49 139 66 hn hn NOUN njb-49 139 67 pcr pcr NOUN njb-49 139 68 product product NOUN njb-49 139 69 ( ( PUNCT njb-49 139 70 1266 1266 NUM njb-49 139 71 bp bp PROPN njb-49 139 72 ) ) PUNCT njb-49 139 73 3000 3000 NUM njb-49 139 74 m m SYM njb-49 139 75 1 1 NUM njb-49 139 76 500 500 NUM njb-49 139 77 1000 1000 NUM njb-49 139 78 850 850 NUM njb-49 139 79 650 650 NUM njb-49 139 80 1650 1650 NUM njb-49 139 81 2000 2000 NUM njb-49 140 1 bp bp PROPN njb-49 140 2 b b PROPN njb-49 140 3 ( ( PUNCT njb-49 140 4 i i NOUN njb-49 140 5 ) ) PUNCT njb-49 140 6 m m VERB njb-49 140 7 1 1 NUM njb-49 140 8 insert insert PROPN njb-49 140 9 f f PROPN njb-49 140 10 - - PUNCT njb-49 140 11 hn hn PROPN njb-49 140 12 ( ( PUNCT njb-49 140 13 1266 1266 NUM njb-49 140 14 bp bp PROPN njb-49 140 15 ) ) PUNCT njb-49 140 16 500 500 NUM njb-49 140 17 650 650 NUM njb-49 140 18 1000 1000 NUM njb-49 140 19 850 850 NUM njb-49 140 20 1650 1650 NUM njb-49 140 21 3000 3000 NUM njb-49 140 22 2000 2000 NUM njb-49 140 23 4000 4000 NUM njb-49 140 24 5000 5000 NUM njb-49 140 25 bp bp PROPN njb-49 140 26 vector vector NOUN njb-49 140 27 pgex-4t-1 pgex-4t-1 PROPN njb-49 140 28 ( ( PUNCT njb-49 140 29 4.9 4.9 NUM njb-49 140 30 kb kb PROPN njb-49 140 31 ) ) PUNCT njb-49 140 32 b b PROPN njb-49 140 33 ( ( PUNCT njb-49 140 34 ii ii PROPN njb-49 140 35 ) ) PUNCT njb-49 140 36 m m VERB njb-49 140 37 1 1 NUM njb-49 140 38 amplified amplify VERB njb-49 140 39 f f PROPN njb-49 140 40 - - PUNCT njb-49 140 41 hc hc NOUN njb-49 140 42 pcr pcr NOUN njb-49 140 43 product product NOUN njb-49 140 44 ( ( PUNCT njb-49 140 45 1263 1263 NUM njb-49 140 46 bp bp PROPN njb-49 140 47 ) ) PUNCT njb-49 140 48 bp bp PROPN njb-49 140 49 100 100 NUM njb-49 140 50 400 400 NUM njb-49 140 51 500 500 NUM njb-49 140 52 3000 3000 NUM njb-49 140 53 2000 2000 NUM njb-49 140 54 5000 5000 NUM njb-49 140 55 1000 1000 NUM njb-49 140 56 300 300 NUM njb-49 140 57 200 200 NUM njb-49 140 58 650 650 NUM njb-49 140 59 850 850 NUM njb-49 140 60 1650 1650 NUM njb-49 140 61 4000 4000 NUM njb-49 140 62 c c NOUN njb-49 140 63 ( ( PUNCT njb-49 140 64 i i NOUN njb-49 140 65 ) ) PUNCT njb-49 140 66 m m VERB njb-49 140 67 1 1 NUM njb-49 140 68 insert insert PROPN njb-49 140 69 f f X njb-49 140 70 - - PUNCT njb-49 140 71 hc hc PROPN njb-49 140 72 ( ( PUNCT njb-49 140 73 1263 1263 NUM njb-49 140 74 bp bp PROPN njb-49 140 75 ) ) PUNCT njb-49 140 76 vector vector NOUN njb-49 140 77 pgex-4t-1 pgex-4t-1 PROPN njb-49 140 78 ( ( PUNCT njb-49 140 79 4.9 4.9 NUM njb-49 140 80 kb kb NUM njb-49 140 81 ) ) PUNCT njb-49 140 82 300 300 NUM njb-49 140 83 400 400 NUM njb-49 140 84 850 850 NUM njb-49 140 85 650 650 NUM njb-49 140 86 1650 1650 NUM njb-49 140 87 3000 3000 NUM njb-49 140 88 2000 2000 NUM njb-49 140 89 4000 4000 NUM njb-49 140 90 5000 5000 NUM njb-49 140 91 500 500 NUM njb-49 140 92 1000 1000 NUM njb-49 140 93 bp bp PROPN njb-49 140 94 c c PROPN njb-49 140 95 ( ( PUNCT njb-49 140 96 ii ii PROPN njb-49 140 97 ) ) PUNCT njb-49 140 98 2000 2000 NUM njb-49 140 99 1000 1000 NUM njb-49 140 100 1650 1650 NUM njb-49 140 101 4000 4000 NUM njb-49 140 102 m m NUM njb-49 140 103 1 1 NUM njb-49 140 104 amplified amplify VERB njb-49 140 105 f f PROPN njb-49 140 106 - - NOUN njb-49 140 107 hcc hcc PROPN njb-49 140 108 pcr pcr PROPN njb-49 140 109 product product NOUN njb-49 140 110 ( ( PUNCT njb-49 140 111 501 501 NUM njb-49 140 112 bp bp PROPN njb-49 140 113 ) ) PUNCT njb-49 140 114 bp bp PROPN njb-49 140 115 100 100 NUM njb-49 140 116 400 400 NUM njb-49 140 117 500 500 NUM njb-49 140 118 300 300 NUM njb-49 140 119 200 200 NUM njb-49 140 120 650 650 NUM njb-49 140 121 850 850 NUM njb-49 140 122 d d X njb-49 140 123 ( ( PUNCT njb-49 140 124 i i NOUN njb-49 140 125 ) ) PUNCT njb-49 140 126 m m VERB njb-49 140 127 1 1 NUM njb-49 140 128 insert insert PROPN njb-49 140 129 f f PROPN njb-49 140 130 - - PUNCT njb-49 140 131 hcc hcc PROPN njb-49 140 132 ( ( PUNCT njb-49 140 133 501 501 NUM njb-49 140 134 bp bp PROPN njb-49 140 135 ) ) PUNCT njb-49 140 136 vector vector NOUN njb-49 140 137 pgex-4t-1 pgex-4t-1 PROPN njb-49 140 138 ( ( PUNCT njb-49 140 139 4.9 4.9 NUM njb-49 140 140 kb kb NOUN njb-49 140 141 ) ) PUNCT njb-49 140 142 d d PROPN njb-49 140 143 ( ( PUNCT njb-49 140 144 ii ii PROPN njb-49 140 145 ) ) PUNCT njb-49 140 146 2000 2000 NUM njb-49 140 147 1000 1000 NUM njb-49 140 148 1650 1650 NUM njb-49 140 149 4000 4000 NUM njb-49 140 150 bp bp PROPN njb-49 140 151 100 100 NUM njb-49 140 152 400 400 NUM njb-49 140 153 500 500 NUM njb-49 140 154 300 300 NUM njb-49 140 155 200 200 NUM njb-49 140 156 650 650 NUM njb-49 140 157 850 850 NUM njb-49 140 158 fig.3 fig.3 PROPN njb-49 140 159 : : PUNCT njb-49 140 160 agarose agarose NOUN njb-49 140 161 gel gel NOUN njb-49 140 162 ( ( PUNCT njb-49 140 163 1 1 NUM njb-49 140 164 % % NOUN njb-49 140 165 ) ) PUNCT njb-49 140 166 electrophoresis electrophoresis NOUN njb-49 140 167 of of ADP njb-49 140 168 pcr pcr NOUN njb-49 140 169 product product NOUN njb-49 140 170 and and CCONJ njb-49 140 171 restriction restriction NOUN njb-49 140 172 digested digest VERB njb-49 140 173 plasmid plasmid NOUN njb-49 140 174 stained stain VERB njb-49 140 175 with with ADP njb-49 140 176 ethidium ethidium NOUN njb-49 140 177 bromide bromide NOUN njb-49 140 178 and and CCONJ njb-49 140 179 visualized visualize VERB njb-49 140 180 by by ADP njb-49 140 181 uv uv PROPN njb-49 140 182 nepal nepal ADJ njb-49 140 183     SPACE njb-49 140 184 journal journal NOUN njb-49 140 185     SPACE njb-49 140 186 of of ADP njb-49 140 187     SPACE njb-49 140 188 biotechnology biotechnology NOUN njb-49 140 189 . . PUNCT njb-49 140 190         SPACE njb-49 141 1 jan jan PROPN njb-49 141 2 . . PROPN njb-49 141 3     SPACE njb-49 141 4 2012 2012 NUM njb-49 141 5 , , PUNCT njb-49 141 6     SPACE njb-49 141 7 vol vol NOUN njb-49 141 8 . . PUNCT njb-49 141 9     SPACE njb-49 141 10 2 2 NUM njb-49 141 11 , , PUNCT njb-49 141 12     SPACE njb-49 141 13 no no INTJ njb-49 141 14 . . PUNCT njb-49 141 15     SPACE njb-49 142 1 1 1 NUM njb-49 142 2 : : PUNCT njb-49 142 3     SPACE njb-49 142 4 1 1 X njb-49 142 5     SPACE njb-49 142 6 – – PUNCT njb-49 142 7     SPACE njb-49 142 8 15 15 NUM njb-49 142 9                                                                                                                                                                                                                                         SPACE njb-49 142 10 biotechnology biotechnology NOUN njb-49 142 11     SPACE njb-49 142 12 society society NOUN njb-49 142 13     SPACE njb-49 142 14 of of ADP njb-49 142 15     SPACE njb-49 142 16 nepal nepal NOUN njb-49 142 17     SPACE njb-49 142 18 ( ( PUNCT njb-49 142 19 bsn bsn PROPN njb-49 142 20 ) ) PUNCT njb-49 142 21 , , PUNCT njb-49 142 22     SPACE njb-49 142 23 all all DET njb-49 142 24     SPACE njb-49 142 25 rights right NOUN njb-49 142 26     SPACE njb-49 142 27 reserved reserve VERB njb-49 142 28     SPACE njb-49 142 29 9 9 NUM njb-49 142 30 iptg iptg NOUN njb-49 142 31 induction induction NOUN njb-49 142 32 was be AUX njb-49 142 33 carried carry VERB njb-49 142 34 out out ADP njb-49 142 35 in in ADP njb-49 142 36 a a DET njb-49 142 37 time time NOUN njb-49 142 38 dependent dependent ADJ njb-49 142 39 manner manner NOUN njb-49 142 40 and and CCONJ njb-49 142 41 sds sds NOUN njb-49 142 42 – – PUNCT njb-49 142 43 page page NOUN njb-49 142 44 ( ( PUNCT njb-49 142 45 12 12 NUM njb-49 142 46 % % NOUN njb-49 142 47 ) ) PUNCT njb-49 142 48 analysis analysis NOUN njb-49 142 49 of of ADP njb-49 142 50 total total ADJ njb-49 142 51 lysate lysate NOUN njb-49 142 52 of of ADP njb-49 142 53 e. e. PROPN njb-49 142 54 coli coli PROPN njb-49 142 55 bl21(pgex-4t-1 bl21(pgex-4t-1 PROPN njb-49 142 56 - - PUNCT njb-49 142 57 f f PROPN njb-49 142 58 - - PUNCT njb-49 142 59 lc lc PROPN njb-49 142 60 ( ( PUNCT njb-49 142 61 a a NOUN njb-49 142 62 ) ) PUNCT njb-49 142 63 and and CCONJ njb-49 142 64 pgex-4t-1 pgex-4t-1 PROPN njb-49 142 65 - - PUNCT njb-49 142 66 f f PROPN njb-49 142 67 - - PUNCT njb-49 142 68 hn hn PROPN njb-49 142 69 ( ( PUNCT njb-49 142 70 b b NOUN njb-49 142 71 ) ) PUNCT njb-49 142 72 with with ADP njb-49 142 73 or or CCONJ njb-49 142 74 without without ADP njb-49 142 75 iptg iptg NOUN njb-49 142 76 induction induction NOUN njb-49 142 77 stained stain VERB njb-49 142 78 with with ADP njb-49 142 79 coomassie coomassie ADJ njb-49 142 80 blue blue NOUN njb-49 142 81 . . PUNCT njb-49 143 1 lane lane PROPN njb-49 143 2 m m NOUN njb-49 143 3 : : PUNCT njb-49 143 4 protein protein NOUN njb-49 143 5 marker marker NOUN njb-49 143 6 ( ( PUNCT njb-49 143 7 bio bio NOUN njb-49 143 8 - - ADJ njb-49 143 9 rad rad ADJ njb-49 143 10 ) ) PUNCT njb-49 143 11 ; ; PUNCT njb-49 143 12 lanes lane NOUN njb-49 143 13 1 1 NUM njb-49 143 14 4 4 NUM njb-49 143 15 : : PUNCT njb-49 143 16 ( ( PUNCT njb-49 143 17 0 0 NUM njb-49 143 18 , , PUNCT njb-49 143 19 2 2 NUM njb-49 143 20 , , PUNCT njb-49 143 21 5 5 NUM njb-49 143 22 , , PUNCT njb-49 143 23 and and CCONJ njb-49 143 24 7 7 NUM njb-49 143 25 h h NOUN njb-49 143 26 for for ADP njb-49 143 27 f f PROPN njb-49 143 28 - - PUNCT njb-49 143 29 lc lc NOUN njb-49 143 30 and and CCONJ njb-49 143 31 0 0 NUM njb-49 143 32 , , PUNCT njb-49 143 33 2 2 NUM njb-49 143 34 , , PUNCT njb-49 143 35 5 5 NUM njb-49 143 36 , , PUNCT njb-49 143 37 and and CCONJ njb-49 143 38 8 8 NUM njb-49 143 39 h h NOUN njb-49 143 40 for for ADP njb-49 143 41 f f PROPN njb-49 143 42 - - NOUN njb-49 143 43 hn hn PROPN njb-49 143 44 ) ) PUNCT njb-49 143 45 after after ADP njb-49 143 46 iptg iptg PROPN njb-49 143 47 induction induction NOUN njb-49 143 48 . . PUNCT njb-49 144 1 lanes lane VERB njb-49 144 2 2 2 NUM njb-49 144 3 4 4 NUM njb-49 144 4 demonstrate demonstrate NOUN njb-49 144 5 expressed express VERB njb-49 144 6 gst gst NOUN njb-49 144 7 - - PUNCT njb-49 144 8 f f PROPN njb-49 144 9 - - PUNCT njb-49 144 10 lc lc NOUN njb-49 144 11 a a NOUN njb-49 144 12 ) ) PUNCT njb-49 144 13 and and CCONJ njb-49 144 14 gst gst PROPN njb-49 144 15 - - PUNCT njb-49 144 16 f f PROPN njb-49 144 17 - - PUNCT njb-49 144 18 hn hn PROPN njb-49 144 19 ( ( PUNCT njb-49 144 20 b b NOUN njb-49 144 21 ) ) PUNCT njb-49 144 22 with with ADP njb-49 144 23 treatment treatment NOUN njb-49 144 24 of of ADP njb-49 144 25 iptg iptg PROPN njb-49 144 26 . . PUNCT njb-49 145 1 kda kda PROPN njb-49 145 2 , , PUNCT njb-49 145 3 protein protein NOUN njb-49 145 4 size size NOUN njb-49 145 5 expressed express VERB njb-49 145 6 in in ADP njb-49 145 7 kilodalton kilodalton PROPN njb-49 145 8 . . PUNCT njb-49 146 1 although although SCONJ njb-49 146 2 both both DET njb-49 146 3 vectors vector NOUN njb-49 146 4 contain contain VERB njb-49 146 5 thrombin thrombin NOUN njb-49 146 6 cleavage cleavage NOUN njb-49 146 7 sites site NOUN njb-49 146 8 and and CCONJ njb-49 146 9 are be AUX njb-49 146 10 ideal ideal ADJ njb-49 146 11 for for ADP njb-49 146 12 production production NOUN njb-49 146 13 of of ADP njb-49 146 14 soluble soluble ADJ njb-49 146 15 proteins protein NOUN njb-49 146 16 , , PUNCT njb-49 146 17 neither neither DET njb-49 146 18 one one NUM njb-49 146 19 was be AUX njb-49 146 20 able able ADJ njb-49 146 21 to to PART njb-49 146 22 alter alter VERB njb-49 146 23 the the DET njb-49 146 24 formation formation NOUN njb-49 146 25 of of ADP njb-49 146 26 inclusion inclusion NOUN njb-49 146 27 bodies body NOUN njb-49 146 28 . . PUNCT njb-49 147 1 proteinrefolding proteinrefolde VERB njb-49 147 2 procedures procedure NOUN njb-49 147 3 were be AUX njb-49 147 4 performed perform VERB njb-49 147 5 using use VERB njb-49 147 6 reducing reduce VERB njb-49 147 7 reagents reagent NOUN njb-49 147 8 including include VERB njb-49 147 9 various various ADJ njb-49 147 10 molar molar NOUN njb-49 147 11 concentrations concentration NOUN njb-49 147 12 of of ADP njb-49 147 13 urea urea NOUN njb-49 147 14 or or CCONJ njb-49 147 15 guanidine guanidine NOUN njb-49 147 16 – – PUNCT njb-49 147 17 hcl hcl PROPN njb-49 147 18 to to PART njb-49 147 19 solubilize solubilize VERB njb-49 147 20 the the DET njb-49 147 21 inclusion inclusion NOUN njb-49 147 22 bodies body NOUN njb-49 147 23 but but CCONJ njb-49 147 24 attempts attempt NOUN njb-49 147 25 were be AUX njb-49 147 26 unsuccessful unsuccessful ADJ njb-49 147 27 . . PUNCT njb-49 148 1 as as ADP njb-49 148 2 a a DET njb-49 148 3 result result NOUN njb-49 148 4 , , PUNCT njb-49 148 5 only only ADV njb-49 148 6 lc lc NOUN njb-49 148 7 and and CCONJ njb-49 148 8 hn hn PROPN njb-49 148 9 domains domain NOUN njb-49 148 10 were be AUX njb-49 148 11 used use VERB njb-49 148 12 for for ADP njb-49 148 13 protein protein NOUN njb-49 148 14 analysis analysis NOUN njb-49 148 15 and and CCONJ njb-49 148 16 collection collection NOUN njb-49 148 17 of of ADP njb-49 148 18 polyclonal polyclonal ADJ njb-49 148 19 serum serum NOUN njb-49 148 20 from from ADP njb-49 148 21 immunized immunized ADJ njb-49 148 22 mice mouse NOUN njb-49 148 23 . . PUNCT njb-49 149 1 gst gst PROPN njb-49 149 2 affinity affinity PROPN njb-49 149 3 chromatography chromatography NOUN njb-49 149 4 was be AUX njb-49 149 5 then then ADV njb-49 149 6 carried carry VERB njb-49 149 7 out out ADP njb-49 149 8 to to PART njb-49 149 9 collect collect VERB njb-49 149 10 gst gst NOUN njb-49 149 11 tagged tag VERB njb-49 149 12 lc lc NOUN njb-49 150 1 and and CCONJ njb-49 151 1 hn hn PROPN njb-49 151 2 . . PUNCT njb-49 152 1 after after ADP njb-49 152 2 gst gst PROPN njb-49 152 3 affinity affinity PROPN njb-49 152 4 chromatography chromatography NOUN njb-49 152 5 and and CCONJ njb-49 152 6 cleavage cleavage NOUN njb-49 152 7 with with ADP njb-49 152 8 thrombin thrombin NOUN njb-49 152 9 protease protease NOUN njb-49 152 10 treatment treatment NOUN njb-49 152 11 , , PUNCT njb-49 152 12 recombinant recombinant ADJ njb-49 152 13 gst gst NOUN njb-49 152 14 free free ADJ njb-49 152 15 f f X njb-49 152 16 - - PUNCT njb-49 152 17 lc lc NOUN njb-49 152 18 and and CCONJ njb-49 152 19 gst gst NOUN njb-49 152 20 free free ADJ njb-49 152 21 f f X njb-49 152 22 - - PROPN njb-49 152 23 hn hn PROPN njb-49 152 24 were be AUX njb-49 152 25 analyzed analyze VERB njb-49 152 26 on on ADP njb-49 152 27 sds sds NOUN njb-49 152 28 - - PUNCT njb-49 152 29 page page NOUN njb-49 152 30 to to PART njb-49 152 31 monitor monitor VERB njb-49 152 32 expected expect VERB njb-49 152 33 protein protein NOUN njb-49 152 34 bands band NOUN njb-49 152 35 . . PUNCT njb-49 153 1 sds sds NOUN njb-49 153 2 - - PUNCT njb-49 153 3 page page NOUN njb-49 153 4 revealed reveal VERB njb-49 153 5 the the DET njb-49 153 6 appropriate appropriate ADJ njb-49 153 7 size size NOUN njb-49 153 8 of of ADP njb-49 153 9 both both DET njb-49 153 10 recombinants recombinant NOUN njb-49 153 11 ; ; PUNCT njb-49 153 12 gst gst PROPN njb-49 153 13 fused fuse VERB njb-49 153 14 75 75 NUM njb-49 153 15 kda kda NOUN njb-49 153 16 and and CCONJ njb-49 153 17 gst gst NOUN njb-49 153 18 free free ADJ njb-49 153 19 50 50 NUM njb-49 153 20 kda kda PROPN njb-49 153 21 ( ( PUNCT njb-49 153 22 fig fig NOUN njb-49 153 23 . . PUNCT njb-49 153 24 5 5 NUM njb-49 153 25 ) ) PUNCT njb-49 153 26 . . PUNCT njb-49 154 1 final final ADJ njb-49 154 2 recombinant recombinant ADJ njb-49 154 3 protein protein NOUN njb-49 154 4 concentrations concentration NOUN njb-49 154 5 were be AUX njb-49 154 6 determined determine VERB njb-49 154 7 to to PART njb-49 154 8 be be AUX njb-49 154 9 1.0 1.0 NUM njb-49 154 10 mg mg NOUN njb-49 154 11 / / SYM njb-49 154 12 l. l. PROPN njb-49 154 13 in in ADP njb-49 154 14 order order NOUN njb-49 154 15 to to PART njb-49 154 16 analyze analyze VERB njb-49 154 17 specificity specificity NOUN njb-49 154 18 of of ADP njb-49 154 19 mice mouse NOUN njb-49 154 20 polyclonal polyclonal ADJ njb-49 154 21 sera sera PROPN njb-49 154 22 against against ADP njb-49 154 23 flc flc PROPN njb-49 154 24 and and CCONJ njb-49 154 25 f f PROPN njb-49 154 26 - - PUNCT njb-49 154 27 hn hn PROPN njb-49 154 28 , , PUNCT njb-49 154 29 mice mouse NOUN njb-49 154 30 were be AUX njb-49 154 31 immunized immunize VERB njb-49 154 32 intraperitoneally intraperitoneally ADV njb-49 154 33 with with ADP njb-49 154 34 antigens antigen NOUN njb-49 154 35 ; ; PUNCT njb-49 154 36 gst gst PROPN njb-49 154 37 free free PROPN njb-49 154 38 flc flc PROPN njb-49 154 39 and and CCONJ njb-49 154 40 f f PROPN njb-49 154 41 - - NOUN njb-49 154 42 hn hn PROPN njb-49 154 43 . . PROPN njb-49 155 1 to to PART njb-49 155 2 analyze analyze VERB njb-49 155 3 the the DET njb-49 155 4 specificity specificity NOUN njb-49 155 5 of of ADP njb-49 155 6 the the DET njb-49 155 7 polyclonal polyclonal ADJ njb-49 155 8 serums serum NOUN njb-49 155 9 against against ADP njb-49 155 10 f f PROPN njb-49 155 11 - - PUNCT njb-49 155 12 lc lc NOUN njb-49 155 13 and and CCONJ njb-49 155 14 fhn fhn PROPN njb-49 155 15 , , PUNCT njb-49 155 16 mouse mouse NOUN njb-49 155 17 polyclonal polyclonal ADJ njb-49 155 18 serum serum NOUN njb-49 155 19 was be AUX njb-49 155 20 collected collect VERB njb-49 155 21 from from ADP njb-49 155 22 the the DET njb-49 155 23 tail tail NOUN njb-49 155 24 vain vain ADJ njb-49 155 25 and and CCONJ njb-49 155 26 tested test VERB njb-49 155 27 by by ADP njb-49 155 28 western western ADJ njb-49 155 29 blotting blotting NOUN njb-49 155 30 and and CCONJ njb-49 155 31 indirect indirect ADJ njb-49 155 32 elisa elisa NOUN njb-49 155 33 ( ( PUNCT njb-49 155 34 both both PRON njb-49 155 35 of of ADP njb-49 155 36 these these DET njb-49 155 37 techniques technique NOUN njb-49 155 38 being be AUX njb-49 155 39 highly highly ADV njb-49 155 40 valuable valuable ADJ njb-49 155 41 in in ADP njb-49 155 42 the the DET njb-49 155 43 detection detection NOUN njb-49 155 44 of of ADP njb-49 155 45 candidate candidate NOUN njb-49 155 46 biomarkers biomarker NOUN njb-49 155 47 ) ) PUNCT njb-49 155 48 . . PUNCT njb-49 156 1 the the DET njb-49 156 2 mouse mouse NOUN njb-49 156 3 anti anti ADJ njb-49 156 4 - - ADJ njb-49 156 5 f f ADJ njb-49 156 6 - - PUNCT njb-49 156 7 lc lc NOUN njb-49 156 8 polyclonal polyclonal ADJ njb-49 156 9 antibodies antibody NOUN njb-49 156 10 were be AUX njb-49 156 11 found find VERB njb-49 156 12 to to PART njb-49 156 13 specifically specifically ADV njb-49 156 14 recognize recognize VERB njb-49 156 15 f f PROPN njb-49 156 16 - - PUNCT njb-49 156 17 lc lc NOUN njb-49 156 18 and and CCONJ njb-49 156 19 bont bont PROPN njb-49 156 20 / / SYM njb-49 156 21 f f PROPN njb-49 156 22 ( ( PUNCT njb-49 156 23 figs fig NOUN njb-49 156 24 . . PUNCT njb-49 157 1 5a 5a NUM njb-49 157 2 and and CCONJ njb-49 157 3 6a 6a NUM njb-49 157 4 ) ) PUNCT njb-49 157 5 . . PUNCT njb-49 158 1 similarly similarly ADV njb-49 158 2 , , PUNCT njb-49 158 3 anti anti ADJ njb-49 158 4 - - ADJ njb-49 158 5 fhn fhn ADJ njb-49 158 6 polyclonal polyclonal ADJ njb-49 158 7 antibodies antibody NOUN njb-49 158 8 demonstrated demonstrate VERB njb-49 158 9 a a DET njb-49 158 10 m m NOUN njb-49 158 11 1 1 NUM njb-49 158 12 2 2 NUM njb-49 158 13 3 3 NUM njb-49 158 14 4 4 NUM njb-49 158 15 kda kda NOUN njb-49 158 16 25 25 NUM njb-49 158 17 75 75 NUM njb-49 158 18 50 50 NUM njb-49 158 19 37 37 NUM njb-49 158 20 100 100 NUM njb-49 158 21 150 150 NUM njb-49 158 22 250 250 NUM njb-49 158 23 15 15 NUM njb-49 158 24 10 10 NUM njb-49 158 25 m m NUM njb-49 158 26 1 1 NUM njb-49 158 27 2 2 NUM njb-49 158 28 3 3 NUM njb-49 158 29 4 4 NUM njb-49 158 30 kda kda NOUN njb-49 158 31 25 25 NUM njb-49 158 32 75 75 NUM njb-49 158 33 50 50 NUM njb-49 158 34 37 37 NUM njb-49 158 35 100 100 NUM njb-49 158 36 150 150 NUM njb-49 158 37 250 250 NUM njb-49 158 38 15 15 NUM njb-49 158 39 gst gst NOUN njb-49 158 40 - - PUNCT njb-49 158 41 f f PROPN njb-49 158 42 - - PUNCT njb-49 158 43 lc lc NOUN njb-49 158 44 gst gst NOUN njb-49 158 45 - - PUNCT njb-49 158 46 f f PROPN njb-49 158 47 - - PUNCT njb-49 158 48 hn hn PROPN njb-49 158 49 sds sds PROPN njb-49 158 50 - - PUNCT njb-49 158 51 page page NOUN njb-49 158 52 ( ( PUNCT njb-49 158 53 12 12 NUM njb-49 158 54 % % NOUN njb-49 158 55 ) ) PUNCT njb-49 158 56 4 4 NUM njb-49 158 57 ( ( PUNCT njb-49 158 58 a a PRON njb-49 158 59 ) ) PUNCT njb-49 158 60 sds sds NOUN njb-49 158 61 - - PUNCT njb-49 158 62 page page NOUN njb-49 158 63 ( ( PUNCT njb-49 158 64 12 12 NUM njb-49 158 65 % % NOUN njb-49 158 66 ) ) PUNCT njb-49 158 67 4 4 NUM njb-49 158 68 ( ( PUNCT njb-49 158 69 b b NOUN njb-49 158 70 ) ) PUNCT njb-49 158 71 fig.4 fig.4 PROPN njb-49 158 72 : : PUNCT njb-49 158 73 induction induction NOUN njb-49 158 74 and and CCONJ njb-49 158 75 expression expression NOUN njb-49 158 76 analysis analysis NOUN njb-49 158 77 of of ADP njb-49 158 78 recombinant recombinant ADJ njb-49 158 79 proteins protein NOUN njb-49 158 80 nepal nepal ADJ njb-49 158 81     SPACE njb-49 158 82 journal journal NOUN njb-49 158 83     SPACE njb-49 158 84 of of ADP njb-49 158 85     SPACE njb-49 158 86 biotechnology biotechnology NOUN njb-49 158 87 . . PUNCT njb-49 158 88         SPACE njb-49 159 1 jan jan PROPN njb-49 159 2 . . PROPN njb-49 159 3     SPACE njb-49 159 4 2012 2012 NUM njb-49 159 5 , , PUNCT njb-49 159 6     SPACE njb-49 159 7 vol vol NOUN njb-49 159 8 . . PUNCT njb-49 159 9     SPACE njb-49 159 10 2 2 NUM njb-49 159 11 , , PUNCT njb-49 159 12     SPACE njb-49 159 13 no no INTJ njb-49 159 14 . . PUNCT njb-49 159 15     SPACE njb-49 160 1 1 1 NUM njb-49 160 2 : : PUNCT njb-49 160 3     SPACE njb-49 160 4 1 1 X njb-49 160 5     SPACE njb-49 160 6 – – PUNCT njb-49 160 7     SPACE njb-49 160 8 15 15 NUM njb-49 160 9                                                                                                                                                                                                                                         SPACE njb-49 160 10 biotechnology biotechnology NOUN njb-49 160 11     SPACE njb-49 160 12 society society NOUN njb-49 160 13     SPACE njb-49 160 14 of of ADP njb-49 160 15     SPACE njb-49 160 16 nepal nepal NOUN njb-49 160 17     SPACE njb-49 160 18 ( ( PUNCT njb-49 160 19 bsn bsn PROPN njb-49 160 20 ) ) PUNCT njb-49 160 21 , , PUNCT njb-49 160 22     SPACE njb-49 160 23 all all DET njb-49 160 24     SPACE njb-49 160 25 rights right NOUN njb-49 160 26     SPACE njb-49 160 27 reserved reserve VERB njb-49 160 28     SPACE njb-49 160 29 10 10 NUM njb-49 160 30 strong strong ADJ njb-49 160 31 affinity affinity NOUN njb-49 160 32 towards towards ADP njb-49 160 33 f f PROPN njb-49 160 34 - - PROPN njb-49 160 35 hn hn NOUN njb-49 160 36 and and CCONJ njb-49 160 37 native native ADJ njb-49 160 38 type type NOUN njb-49 160 39 f f PROPN njb-49 160 40 toxin toxin PROPN njb-49 160 41 ( ( PUNCT njb-49 160 42 figs fig NOUN njb-49 160 43 . . PUNCT njb-49 161 1 5b 5b NOUN njb-49 161 2 and and CCONJ njb-49 161 3 6b 6b NUM njb-49 161 4 ) ) PUNCT njb-49 161 5 . . PUNCT njb-49 162 1 however however ADV njb-49 162 2 , , PUNCT njb-49 162 3 neither neither CCONJ njb-49 162 4 anti anti ADJ njb-49 162 5 - - ADJ njb-49 162 6 f f ADJ njb-49 162 7 - - PUNCT njb-49 162 8 lc lc NOUN njb-49 162 9 nor nor CCONJ njb-49 162 10 anti anti ADJ njb-49 162 11 - - ADJ njb-49 162 12 f f ADJ njb-49 162 13 - - PUNCT njb-49 162 14 hn hn PROPN njb-49 162 15 polyclonal polyclonal ADJ njb-49 162 16 antibodies antibody NOUN njb-49 162 17 were be AUX njb-49 162 18 able able ADJ njb-49 162 19 to to PART njb-49 162 20 detect detect VERB njb-49 162 21 the the DET njb-49 162 22 hc hc PROPN njb-49 162 23 domain domain NOUN njb-49 162 24 ( ( PUNCT njb-49 162 25 fig fig NOUN njb-49 162 26 . . PROPN njb-49 162 27 5 5 NUM njb-49 162 28 ) ) PUNCT njb-49 162 29 , , PUNCT njb-49 162 30 validating validate VERB njb-49 162 31 the the DET njb-49 162 32 specificity specificity NOUN njb-49 162 33 of of ADP njb-49 162 34 the the DET njb-49 162 35 polyclonal polyclonal ADJ njb-49 162 36 serums serum NOUN njb-49 162 37 towards towards ADP njb-49 162 38 their their PRON njb-49 162 39 respective respective ADJ njb-49 162 40 region region NOUN njb-49 162 41 of of ADP njb-49 162 42 bont bont PROPN njb-49 162 43 / / SYM njb-49 162 44 f. f. PROPN njb-49 162 45 the the DET njb-49 162 46 designated designated ADJ njb-49 162 47 recombinant recombinant ADJ njb-49 162 48 domains domain NOUN njb-49 162 49 have have VERB njb-49 162 50 native native ADJ njb-49 162 51 structure structure NOUN njb-49 162 52 as as ADP njb-49 162 53 holotoxin holotoxin PROPN njb-49 162 54 and and CCONJ njb-49 162 55 it it PRON njb-49 162 56 was be AUX njb-49 162 57 not not PART njb-49 162 58 unexpected unexpected ADJ njb-49 162 59 for for SCONJ njb-49 162 60 them they PRON njb-49 162 61 to to PART njb-49 162 62 produce produce VERB njb-49 162 63 anti anti ADJ njb-49 162 64 - - NOUN njb-49 162 65 sera sera NOUN njb-49 162 66 against against ADP njb-49 162 67 native native ADJ njb-49 162 68 toxin toxin NOUN njb-49 162 69 ( ( PUNCT njb-49 162 70 bont bont PROPN njb-49 162 71 / / SYM njb-49 162 72 f f NUM njb-49 162 73 ) ) PUNCT njb-49 162 74 . . PUNCT njb-49 163 1 mouse mouse NOUN njb-49 163 2 showing show VERB njb-49 163 3 highest high ADJ njb-49 163 4 immunoreactivity immunoreactivity NOUN njb-49 163 5 was be AUX njb-49 163 6 then then ADV njb-49 163 7 sacrificed sacrifice VERB njb-49 163 8 in in ADP njb-49 163 9 attempt attempt NOUN njb-49 163 10 to to PART njb-49 163 11 isolate isolate VERB njb-49 163 12 a a DET njb-49 163 13 monoclonal monoclonal NOUN njb-49 163 14 antibody antibody NOUN njb-49 163 15 specific specific ADJ njb-49 163 16 to to ADP njb-49 163 17 f f PROPN njb-49 163 18 - - PUNCT njb-49 163 19 lc lc NOUN njb-49 163 20 or or CCONJ njb-49 163 21 fhn fhn NOUN njb-49 163 22 domains domain NOUN njb-49 163 23 . . PUNCT njb-49 164 1 this this PRON njb-49 164 2 could could AUX njb-49 164 3 allow allow VERB njb-49 164 4 further further ADJ njb-49 164 5 research research NOUN njb-49 164 6 into into ADP njb-49 164 7 the the DET njb-49 164 8 development development NOUN njb-49 164 9 of of ADP njb-49 164 10 a a DET njb-49 164 11 highly highly ADV njb-49 164 12 effective effective ADJ njb-49 164 13 neutralizing neutralize VERB njb-49 164 14 monoclonal monoclonal NOUN njb-49 164 15 antibody antibody NOUN njb-49 164 16 for for ADP njb-49 164 17 a a DET njb-49 164 18 subunit subunit NOUN njb-49 164 19 vaccine vaccine NOUN njb-49 164 20 and/or and/or CCONJ njb-49 164 21 passive passive ADJ njb-49 164 22 immunotherapy immunotherapy NOUN njb-49 164 23 against against ADP njb-49 164 24 bont bont PROPN njb-49 164 25 / / SYM njb-49 164 26 f. f. PROPN njb-49 164 27 ( ( PUNCT njb-49 164 28 i i NOUN njb-49 164 29 ) ) PUNCT njb-49 164 30 sds sds PROPN njb-49 164 31 – – PUNCT njb-49 164 32 page page NOUN njb-49 164 33 of of ADP njb-49 164 34 purified purify VERB njb-49 164 35 gst gst NOUN njb-49 164 36 f f PROPN njb-49 164 37 - - PUNCT njb-49 164 38 lc lc NOUN njb-49 164 39 and and CCONJ njb-49 164 40 gst gst NOUN njb-49 164 41 free free ADJ njb-49 164 42 f f X njb-49 164 43 - - PUNCT njb-49 164 44 lc lc PROPN njb-49 164 45 ( ( PUNCT njb-49 164 46 a a X njb-49 164 47 ) ) PUNCT njb-49 164 48 purified purify VERB njb-49 164 49 gst gst NOUN njb-49 164 50 f f PROPN njb-49 164 51 - - PUNCT njb-49 164 52 hn hn PROPN njb-49 164 53 and and CCONJ njb-49 164 54 gst gst X njb-49 164 55 free free ADJ njb-49 164 56 f f X njb-49 164 57 - - PUNCT njb-49 164 58 hn hn PROPN njb-49 164 59 ( ( PUNCT njb-49 164 60 b b NOUN njb-49 164 61 ) ) PUNCT njb-49 164 62 by by ADP njb-49 164 63 single single ADJ njb-49 164 64 step step NOUN njb-49 164 65 gst gst PROPN njb-49 164 66 affinity affinity NOUN njb-49 164 67 chromatography chromatography NOUN njb-49 164 68 separated separate VERB njb-49 164 69 on on ADP njb-49 164 70 a a DET njb-49 164 71 12 12 NUM njb-49 164 72 % % NOUN njb-49 164 73 gel gel NOUN njb-49 164 74 and and CCONJ njb-49 164 75 visualized visualize VERB njb-49 164 76 by by ADP njb-49 164 77 coomassie coomassie ADJ njb-49 164 78 blue blue ADJ njb-49 164 79 stain stain NOUN njb-49 164 80 ; ; PUNCT njb-49 164 81 ( ( PUNCT njb-49 164 82 ii ii X njb-49 164 83 ) ) PUNCT njb-49 164 84 western western ADJ njb-49 164 85 blot blot NOUN njb-49 164 86 analysis analysis NOUN njb-49 164 87 performed perform VERB njb-49 164 88 by by ADP njb-49 164 89 raising raise VERB njb-49 164 90 anti anti ADJ njb-49 164 91 - - ADJ njb-49 164 92 f f ADJ njb-49 164 93 - - PUNCT njb-49 164 94 lc lc PROPN njb-49 164 95 ( ( PUNCT njb-49 164 96 a a NOUN njb-49 164 97 ) ) PUNCT njb-49 164 98 and and CCONJ njb-49 164 99 anti anti ADJ njb-49 164 100 - - ADJ njb-49 164 101 f f ADJ njb-49 164 102 - - PUNCT njb-49 164 103 hn hn PROPN njb-49 164 104 ( ( PUNCT njb-49 164 105 b b PROPN njb-49 164 106 ) ) PUNCT njb-49 164 107 mouse mouse NOUN njb-49 164 108 polyclonal polyclonal ADJ njb-49 164 109 antibody antibody NOUN njb-49 164 110 . . PUNCT njb-49 165 1 lane lane NOUN njb-49 165 2 m m PROPN njb-49 165 3 : : PUNCT njb-49 165 4 protein protein NOUN njb-49 165 5 marker marker NOUN njb-49 165 6 ( ( PUNCT njb-49 165 7 bio bio NOUN njb-49 165 8 - - ADJ njb-49 165 9 rad rad ADJ njb-49 165 10 ) ) PUNCT njb-49 165 11 ; ; PUNCT njb-49 165 12 lane lane NOUN njb-49 165 13 1 1 NUM njb-49 165 14 , , PUNCT njb-49 165 15 gst gst PROPN njb-49 165 16 - - PUNCT njb-49 165 17 f f PROPN njb-49 165 18 - - PUNCT njb-49 165 19 lc lc NOUN njb-49 165 20 or or CCONJ njb-49 165 21 hn hn ADJ njb-49 165 22 ; ; PUNCT njb-49 165 23 lane lane NOUN njb-49 165 24 2 2 NUM njb-49 165 25 , , PUNCT njb-49 165 26 gst gst NOUN njb-49 165 27 free free ADJ njb-49 165 28 - - PUNCT njb-49 165 29 f f NOUN njb-49 165 30 - - PUNCT njb-49 165 31 lc lc NOUN njb-49 165 32 or or CCONJ njb-49 165 33 hn hn ADJ njb-49 165 34 ; ; PUNCT njb-49 165 35 and and CCONJ njb-49 165 36 lane lane NOUN njb-49 165 37 3 3 NUM njb-49 165 38 , , PUNCT njb-49 165 39 unpurified unpurified ADJ njb-49 165 40 gst gst NOUN njb-49 165 41 - - PUNCT njb-49 165 42 f f PROPN njb-49 165 43 - - PUNCT njb-49 165 44 hc hc NOUN njb-49 165 45 . . PUNCT njb-49 166 1 fig.5 fig.5 NOUN njb-49 166 2 : : PUNCT njb-49 166 3 purification purification NOUN njb-49 166 4 and and CCONJ njb-49 166 5 characterization characterization NOUN njb-49 166 6 of of ADP njb-49 166 7 recombinant recombinant ADJ njb-49 166 8 bont bont ADJ njb-49 166 9 / / SYM njb-49 166 10 f f PROPN njb-49 166 11 domain domain NOUN njb-49 166 12 proteins protein NOUN njb-49 166 13 sds sds NOUN njb-49 166 14 - - PUNCT njb-49 166 15 page page NOUN njb-49 166 16 5b 5b NUM njb-49 166 17 ( ( PUNCT njb-49 166 18 i i NOUN njb-49 166 19 ) ) PUNCT njb-49 166 20 m m VERB njb-49 166 21 1 1 NUM njb-49 166 22 2 2 NUM njb-49 166 23 3 3 NUM njb-49 166 24 m m NUM njb-49 166 25 1 1 NUM njb-49 166 26 2 2 NUM njb-49 166 27 3 3 NUM njb-49 166 28 kda kda NOUN njb-49 166 29 25 25 NUM njb-49 166 30 75 75 NUM njb-49 166 31 50 50 NUM njb-49 166 32 37 37 NUM njb-49 166 33 100 100 NUM njb-49 166 34 150 150 NUM njb-49 166 35 250 250 NUM njb-49 166 36 sds sds NOUN njb-49 166 37 - - PUNCT njb-49 166 38 page page NOUN njb-49 166 39 5a 5a NUM njb-49 167 1 ( ( PUNCT njb-49 167 2 i i NOUN njb-49 167 3 ) ) PUNCT njb-49 167 4 kda kda VERB njb-49 167 5 25 25 NUM njb-49 167 6 75 75 NUM njb-49 167 7 50 50 NUM njb-49 167 8 37 37 NUM njb-49 167 9 100 100 NUM njb-49 167 10 150 150 NUM njb-49 167 11 250 250 NUM njb-49 167 12 western western ADJ njb-49 167 13 blot blot NOUN njb-49 167 14 5a 5a NUM njb-49 167 15 ( ( PUNCT njb-49 167 16 ii ii PROPN njb-49 167 17 ) ) PUNCT njb-49 167 18 gst gst PROPN njb-49 167 19 - - PUNCT njb-49 167 20 f f PROPN njb-49 167 21 - - PUNCT njb-49 167 22 lc lc NOUN njb-49 167 23 gst gst NOUN njb-49 167 24 free free ADJ njb-49 167 25 - - PUNCT njb-49 167 26 f f NOUN njb-49 167 27 - - PUNCT njb-49 167 28 lc lc NOUN njb-49 167 29 25 25 NUM njb-49 167 30 75 75 NUM njb-49 167 31 50 50 NUM njb-49 167 32 37 37 NUM njb-49 167 33 100 100 NUM njb-49 167 34 150 150 NUM njb-49 167 35 250 250 NUM njb-49 167 36 m m NUM njb-49 167 37 1 1 NUM njb-49 167 38 2 2 NUM njb-49 167 39 3 3 NUM njb-49 167 40 m m NUM njb-49 167 41 1 1 NUM njb-49 167 42 2 2 NUM njb-49 167 43 3 3 NUM njb-49 167 44 kda kda PROPN njb-49 167 45 kda kda PROPN njb-49 167 46 western western ADJ njb-49 167 47 blot blot NOUN njb-49 167 48 5b 5b NUM njb-49 167 49 ( ( PUNCT njb-49 167 50 ii ii PROPN njb-49 167 51 ) ) PUNCT njb-49 167 52 25 25 NUM njb-49 167 53 75 75 NUM njb-49 167 54 50 50 NUM njb-49 167 55 37 37 NUM njb-49 167 56 100 100 NUM njb-49 167 57 150 150 NUM njb-49 167 58 250 250 NUM njb-49 167 59 gst gst NOUN njb-49 167 60 - - PUNCT njb-49 167 61 f f PROPN njb-49 167 62 - - PUNCT njb-49 167 63 hn hn PROPN njb-49 167 64 gst gst NOUN njb-49 167 65 free free ADJ njb-49 167 66 - - PUNCT njb-49 167 67 f f NOUN njb-49 167 68 - - PUNCT njb-49 167 69 hn hn PROPN njb-49 167 70 nepal nepal ADJ njb-49 167 71     SPACE njb-49 167 72 journal journal NOUN njb-49 167 73     SPACE njb-49 167 74 of of ADP njb-49 167 75     SPACE njb-49 167 76 biotechnology biotechnology NOUN njb-49 167 77 . . PUNCT njb-49 167 78         SPACE njb-49 168 1 jan jan PROPN njb-49 168 2 . . PROPN njb-49 168 3     SPACE njb-49 168 4 2012 2012 NUM njb-49 168 5 , , PUNCT njb-49 168 6     SPACE njb-49 168 7 vol vol NOUN njb-49 168 8 . . PUNCT njb-49 168 9     SPACE njb-49 168 10 2 2 NUM njb-49 168 11 , , PUNCT njb-49 168 12     SPACE njb-49 168 13 no no INTJ njb-49 168 14 . . PUNCT njb-49 168 15     SPACE njb-49 169 1 1 1 NUM njb-49 169 2 : : PUNCT njb-49 169 3     SPACE njb-49 169 4 1 1 X njb-49 169 5     SPACE njb-49 169 6 – – PUNCT njb-49 169 7     SPACE njb-49 169 8 15 15 NUM njb-49 169 9                                                                                                                                                                                                                                         SPACE njb-49 169 10 biotechnology biotechnology NOUN njb-49 169 11     SPACE njb-49 169 12 society society NOUN njb-49 169 13     SPACE njb-49 169 14 of of ADP njb-49 169 15     SPACE njb-49 169 16 nepal nepal NOUN njb-49 169 17     SPACE njb-49 169 18 ( ( PUNCT njb-49 169 19 bsn bsn PROPN njb-49 169 20 ) ) PUNCT njb-49 169 21 , , PUNCT njb-49 169 22     SPACE njb-49 169 23 all all DET njb-49 169 24     SPACE njb-49 169 25 rights right NOUN njb-49 169 26     SPACE njb-49 169 27 reserved reserve VERB njb-49 169 28     SPACE njb-49 169 29 11 11 NUM njb-49 169 30 bar bar NOUN njb-49 169 31 graph graph NOUN njb-49 169 32 values value NOUN njb-49 169 33 represent represent VERB njb-49 169 34 interaction interaction NOUN njb-49 169 35 of of ADP njb-49 169 36 anti anti ADJ njb-49 169 37 f f PROPN njb-49 169 38 - - PUNCT njb-49 169 39 lc lc PROPN njb-49 169 40 ( ( PUNCT njb-49 169 41 a a NOUN njb-49 169 42 ) ) PUNCT njb-49 169 43 and and CCONJ njb-49 169 44 anti anti ADJ njb-49 169 45 - - ADJ njb-49 169 46 f f ADJ njb-49 169 47 - - PUNCT njb-49 169 48 hn hn PROPN njb-49 169 49 ( ( PUNCT njb-49 169 50 b b PROPN njb-49 169 51 ) ) PUNCT njb-49 169 52 mouse mouse NOUN njb-49 169 53 polyclonal polyclonal ADJ njb-49 169 54 serum serum NOUN njb-49 169 55 antibody antibody NOUN njb-49 169 56 with with ADP njb-49 169 57 : : PUNCT njb-49 169 58 pc pc PROPN njb-49 169 59 = = X njb-49 169 60 bont bont NOUN njb-49 169 61 / / SYM njb-49 169 62 f f PROPN njb-49 169 63 as as ADP njb-49 169 64 a a DET njb-49 169 65 coating coating NOUN njb-49 169 66 antigen antigen NOUN njb-49 169 67 ; ; PUNCT njb-49 169 68 s. s. PROPN njb-49 169 69 1 1 NUM njb-49 169 70 3 3 NUM njb-49 169 71 = = NOUN njb-49 169 72 samples sample NOUN njb-49 169 73 of of ADP njb-49 169 74 gst gst X njb-49 169 75 free free ADJ njb-49 169 76 f f X njb-49 169 77 - - PUNCT njb-49 169 78 lc lc PROPN njb-49 169 79 ( ( PUNCT njb-49 169 80 a a NOUN njb-49 169 81 ) ) PUNCT njb-49 169 82 and and CCONJ njb-49 169 83 gst gst NOUN njb-49 169 84 free free ADJ njb-49 169 85 f f X njb-49 169 86 - - PUNCT njb-49 169 87 hn hn PROPN njb-49 169 88 ( ( PUNCT njb-49 169 89 b b PROPN njb-49 169 90 ) ) PUNCT njb-49 169 91 coating coat VERB njb-49 169 92 antigens antigen NOUN njb-49 169 93 respectively respectively ADV njb-49 169 94 & & CCONJ njb-49 169 95 interaction interaction NOUN njb-49 169 96 of of ADP njb-49 169 97 non non ADJ njb-49 169 98 - - ADJ njb-49 169 99 immunized immunized ADJ njb-49 169 100 mouse mouse NOUN njb-49 169 101 serum serum NOUN njb-49 169 102 with with ADP njb-49 169 103 : : PUNCT njb-49 169 104 nc= nc= ADJ njb-49 169 105 gst gst NOUN njb-49 169 106 free free ADJ njb-49 169 107 f f X njb-49 169 108 - - PUNCT njb-49 169 109 lc lc PROPN njb-49 169 110 ( ( PUNCT njb-49 169 111 a a NOUN njb-49 169 112 ) ) PUNCT njb-49 169 113 and and CCONJ njb-49 169 114 gst gst NOUN njb-49 169 115 free free ADJ njb-49 169 116 f f X njb-49 169 117 - - PUNCT njb-49 169 118 hn hn PROPN njb-49 169 119 ( ( PUNCT njb-49 169 120 b b NOUN njb-49 169 121 ) ) PUNCT njb-49 169 122 used use VERB njb-49 169 123 as as ADP njb-49 169 124 coating coat VERB njb-49 169 125 antigens antigen NOUN njb-49 169 126 respectively respectively ADV njb-49 169 127 . . PUNCT njb-49 170 1 mouse mouse NOUN njb-49 170 2 polyclonal polyclonal ADJ njb-49 170 3 serum serum NOUN njb-49 170 4 was be AUX njb-49 170 5 harvested harvest VERB njb-49 170 6 from from ADP njb-49 170 7 three three NUM njb-49 170 8 different different ADJ njb-49 170 9 mice mouse NOUN njb-49 170 10 ( ( PUNCT njb-49 170 11 from from ADP njb-49 170 12 different different ADJ njb-49 170 13 batches batch NOUN njb-49 170 14 ) ) PUNCT njb-49 170 15 immunized immunize VERB njb-49 170 16 with with ADP njb-49 170 17 gst gst X njb-49 170 18 free free ADJ njb-49 170 19 f f X njb-49 170 20 - - PUNCT njb-49 170 21 lc lc NOUN njb-49 170 22 or or CCONJ njb-49 170 23 gst gst NOUN njb-49 170 24 free free ADJ njb-49 170 25 f f X njb-49 170 26 - - PUNCT njb-49 170 27 hn hn PROPN njb-49 170 28 . . PUNCT njb-49 171 1 pcpositive pcpositive ADJ njb-49 171 2 control control NOUN njb-49 171 3 , , PUNCT njb-49 171 4 ncnegative ncnegative ADJ njb-49 171 5 control control NOUN njb-49 171 6 . . PUNCT njb-49 172 1 n n NOUN njb-49 172 2 = = SYM njb-49 172 3 3 3 NUM njb-49 172 4 wells well NOUN njb-49 172 5 , , PUNCT njb-49 172 6 two two NUM njb-49 172 7 - - PUNCT njb-49 172 8 tailed tail VERB njb-49 172 9 students student NOUN njb-49 172 10 t t PROPN njb-49 172 11 - - PUNCT njb-49 172 12 test test PROPN njb-49 172 13 , , PUNCT njb-49 172 14 s. s. PROPN njb-49 172 15 1 1 NUM njb-49 172 16 3 3 NUM njb-49 172 17 compared compare VERB njb-49 172 18 to to ADP njb-49 172 19 nc nc PROPN njb-49 172 20 , , PUNCT njb-49 172 21 p p X njb-49 172 22 < < X njb-49 172 23 0.05 0.05 NUM njb-49 172 24 . . PUNCT njb-49 173 1 attempts attempt NOUN njb-49 173 2 were be AUX njb-49 173 3 unsuccessful unsuccessful ADJ njb-49 173 4 in in ADP njb-49 173 5 isolating isolate VERB njb-49 173 6 a a DET njb-49 173 7 monoclonal monoclonal NOUN njb-49 173 8 antibody antibody NOUN njb-49 173 9 specific specific ADJ njb-49 173 10 towards towards ADP njb-49 173 11 our our PRON njb-49 173 12 constructed construct VERB njb-49 173 13 bont bont ADJ njb-49 173 14 / / SYM njb-49 173 15 f f PROPN njb-49 173 16 domains domain NOUN njb-49 173 17 ( ( PUNCT njb-49 173 18 data datum NOUN njb-49 173 19 not not PART njb-49 173 20 shown show VERB njb-49 173 21 ) ) PUNCT njb-49 173 22 . . PUNCT njb-49 174 1 however however ADV njb-49 174 2 , , PUNCT njb-49 174 3 we we PRON njb-49 174 4 have have AUX njb-49 174 5 successfully successfully ADV njb-49 174 6 isolated isolate VERB njb-49 174 7 a a DET njb-49 174 8 mouse mouse NOUN njb-49 174 9 monoclonal monoclonal NOUN njb-49 174 10 antibody antibody NOUN njb-49 174 11 that that PRON njb-49 174 12 demonstrates demonstrate VERB njb-49 174 13 neutralizing neutralize VERB njb-49 174 14 activity activity NOUN njb-49 174 15 against against ADP njb-49 174 16 native native PROPN njb-49 174 17 bont bont PROPN njb-49 174 18 / / SYM njb-49 174 19 f f PROPN njb-49 174 20 toxin toxin NOUN njb-49 174 21 resulting result VERB njb-49 174 22 from from ADP njb-49 174 23 bont bont PROPN njb-49 174 24 / / SYM njb-49 174 25 f f PROPN njb-49 174 26 toxiod toxiod NOUN njb-49 174 27 in in ADP njb-49 174 28 vivo vivo ADJ njb-49 174 29 immunization immunization NOUN njb-49 174 30 and and CCONJ njb-49 174 31 hybridoma hybridoma NOUN njb-49 174 32 technology technology NOUN njb-49 175 1 [ [ PUNCT njb-49 175 2 38 38 NUM njb-49 175 3 ] ] PUNCT njb-49 175 4 . . PUNCT njb-49 176 1 conclusion conclusion NOUN njb-49 176 2 this this DET njb-49 176 3 paper paper NOUN njb-49 176 4 describes describe VERB njb-49 176 5 molecular molecular ADJ njb-49 176 6 and and CCONJ njb-49 176 7 immunological immunological ADJ njb-49 176 8 studies study NOUN njb-49 176 9 on on ADP njb-49 176 10 c. c. PROPN njb-49 176 11 botulinum botulinum PROPN njb-49 176 12 neurotoxin neurotoxin NOUN njb-49 176 13 type type VERB njb-49 176 14 f f PROPN njb-49 176 15 domains domain NOUN njb-49 176 16 . . PUNCT njb-49 177 1 synthetic synthetic ADJ njb-49 177 2 genes gene NOUN njb-49 177 3 encoding encode VERB njb-49 177 4 bont bont PROPN njb-49 177 5 / / SYM njb-49 177 6 f f PROPN njb-49 177 7 partial partial ADJ njb-49 177 8 fragments fragment NOUN njb-49 177 9 : : PUNCT njb-49 177 10 catalytic catalytic ADJ njb-49 177 11 domain domain NOUN njb-49 177 12 ( ( PUNCT njb-49 177 13 rf rf NOUN njb-49 177 14 - - PUNCT njb-49 177 15 lc lc NOUN njb-49 177 16 ) ) PUNCT njb-49 177 17 , , PUNCT njb-49 177 18 translocation translocation NOUN njb-49 177 19 domain domain NOUN njb-49 177 20 ( ( PUNCT njb-49 177 21 rfhn rfhn PROPN njb-49 177 22 ) ) PUNCT njb-49 177 23 , , PUNCT njb-49 177 24 receptor receptor NOUN njb-49 177 25 binding bind VERB njb-49 177 26 domain domain NOUN njb-49 177 27 ( ( PUNCT njb-49 177 28 rf rf NOUN njb-49 177 29 - - PUNCT njb-49 177 30 hc hc NOUN njb-49 177 31 ) ) PUNCT njb-49 177 32 and and CCONJ njb-49 178 1 quarter quarter ADP njb-49 178 2 part part NOUN njb-49 178 3 of of ADP njb-49 178 4 hc hc PROPN njb-49 178 5 ( ( PUNCT njb-49 178 6 rf rf NOUN njb-49 178 7 - - NOUN njb-49 178 8 hcc hcc PROPN njb-49 178 9 ) ) PUNCT njb-49 178 10 were be AUX njb-49 178 11 designed design VERB njb-49 178 12 and and CCONJ njb-49 178 13 cloned clone VERB njb-49 178 14 into into ADP njb-49 178 15 e. e. PROPN njb-49 178 16 coli coli PROPN njb-49 178 17 . . PUNCT njb-49 179 1 additionally additionally ADV njb-49 179 2 , , PUNCT njb-49 179 3 expression expression NOUN njb-49 179 4 , , PUNCT njb-49 179 5 purification purification NOUN njb-49 179 6 , , PUNCT njb-49 179 7 and and CCONJ njb-49 179 8 immunoreactivites immunoreactivite NOUN njb-49 179 9 were be AUX njb-49 179 10 analyzed analyze VERB njb-49 179 11 by by ADP njb-49 179 12 western western ADJ njb-49 179 13 blotting blotting NOUN njb-49 179 14 and and CCONJ njb-49 179 15 indirect indirect ADJ njb-49 179 16 elisa elisa NOUN njb-49 179 17 for for ADP njb-49 179 18 lc lc NOUN njb-49 179 19 and and CCONJ njb-49 179 20 hn hn PROPN njb-49 179 21 domains domain NOUN njb-49 179 22 . . PUNCT njb-49 180 1 the the DET njb-49 180 2 use use NOUN njb-49 180 3 of of ADP njb-49 180 4 gst gst NOUN njb-49 180 5 tagged tag VERB njb-49 180 6 recombinant recombinant ADJ njb-49 180 7 protein protein NOUN njb-49 180 8 technology technology NOUN njb-49 180 9 was be AUX njb-49 180 10 chosen choose VERB njb-49 180 11 in in ADP njb-49 180 12 order order NOUN njb-49 180 13 to to PART njb-49 180 14 optimize optimize VERB njb-49 180 15 detection detection NOUN njb-49 180 16 and and CCONJ njb-49 180 17 acquisition acquisition NOUN njb-49 180 18 of of ADP njb-49 180 19 high high ADJ njb-49 180 20 purity purity NOUN njb-49 180 21 , , PUNCT njb-49 180 22 stable stable ADJ njb-49 180 23 , , PUNCT njb-49 180 24 and and CCONJ njb-49 180 25 soluble soluble ADJ njb-49 180 26 proteins protein NOUN njb-49 180 27 through through ADP njb-49 180 28 use use NOUN njb-49 180 29 of of ADP njb-49 180 30 single single ADJ njb-49 180 31 step step NOUN njb-49 180 32 affinity affinity NOUN njb-49 180 33 chromatography chromatography NOUN njb-49 180 34 . . PUNCT njb-49 181 1 although although SCONJ njb-49 181 2 host host NOUN njb-49 181 3 strains strain NOUN njb-49 181 4 , , PUNCT njb-49 181 5 vector vector NOUN njb-49 181 6 types type NOUN njb-49 181 7 , , PUNCT njb-49 181 8 and and CCONJ njb-49 181 9 induction induction NOUN njb-49 181 10 conditions condition NOUN njb-49 181 11 for for ADP njb-49 181 12 expression expression NOUN njb-49 181 13 were be AUX njb-49 181 14 optimized optimize VERB njb-49 181 15 to to PART njb-49 181 16 recover recover VERB njb-49 181 17 recombinant recombinant ADJ njb-49 181 18 proteins protein NOUN njb-49 181 19 in in ADP njb-49 181 20 the the DET njb-49 181 21 soluble soluble ADJ njb-49 181 22 fraction fraction NOUN njb-49 181 23 , , PUNCT njb-49 181 24 we we PRON njb-49 181 25 were be AUX njb-49 181 26 unable unable ADJ njb-49 181 27 to to PART njb-49 181 28 purify purify VERB njb-49 181 29 hc hc PROPN njb-49 181 30 and and CCONJ njb-49 181 31 hcc hcc PROPN njb-49 181 32 . . PUNCT njb-49 182 1 immunoreactivity immunoreactivity NOUN njb-49 182 2 was be AUX njb-49 182 3 accessed access VERB njb-49 182 4 through through ADP njb-49 182 5 gst gst X njb-49 182 6 free free ADJ njb-49 182 7 f f X njb-49 182 8 - - PUNCT njb-49 182 9 lc lc NOUN njb-49 182 10 and and CCONJ njb-49 182 11 f f PROPN njb-49 182 12 - - NOUN njb-49 182 13 hn hn NOUN njb-49 182 14 through through ADP njb-49 182 15 in in ADP njb-49 182 16 vivo vivo ADJ njb-49 182 17 immunization immunization NOUN njb-49 182 18 and and CCONJ njb-49 182 19 polyclonal polyclonal ADJ njb-49 182 20 serum serum NOUN njb-49 182 21 antibody antibody NOUN njb-49 182 22 collection collection NOUN njb-49 182 23 , , PUNCT njb-49 182 24 proceeded proceed VERB njb-49 182 25 by by ADP njb-49 182 26 western western ADJ njb-49 182 27 blotting blotting NOUN njb-49 182 28 and and CCONJ njb-49 182 29 indirect indirect ADJ njb-49 182 30 elisa elisa NOUN njb-49 182 31 . . PUNCT njb-49 183 1 the the DET njb-49 183 2 anti anti ADJ njb-49 183 3 - - ADJ njb-49 183 4 f f ADJ njb-49 183 5 - - PUNCT njb-49 183 6 lc lc NOUN njb-49 183 7 and and CCONJ njb-49 183 8 anti anti ADJ njb-49 183 9 - - ADJ njb-49 183 10 f f ADJ njb-49 183 11 - - PUNCT njb-49 183 12 hn hn PROPN njb-49 183 13 mouse mouse PROPN njb-49 183 14 6 6 NUM njb-49 183 15 ( ( PUNCT njb-49 183 16 a a PRON njb-49 183 17 ) ) PUNCT njb-49 183 18 6 6 NUM njb-49 183 19 ( ( PUNCT njb-49 183 20 b b NOUN njb-49 183 21 ) ) PUNCT njb-49 183 22 fig.6 fig.6 PROPN njb-49 183 23 : : PUNCT njb-49 183 24 immunoreactivity immunoreactivity NOUN njb-49 183 25 analysis analysis NOUN njb-49 183 26 by by ADP njb-49 183 27 indirect indirect ADJ njb-49 183 28 elisa elisa PROPN njb-49 183 29 nepal nepal ADJ njb-49 183 30     SPACE njb-49 183 31 journal journal NOUN njb-49 183 32     SPACE njb-49 183 33 of of ADP njb-49 183 34     SPACE njb-49 183 35 biotechnology biotechnology NOUN njb-49 183 36 . . PUNCT njb-49 183 37         SPACE njb-49 184 1 jan jan PROPN njb-49 184 2 . . PROPN njb-49 184 3     SPACE njb-49 184 4 2012 2012 NUM njb-49 184 5 , , PUNCT njb-49 184 6     SPACE njb-49 184 7 vol vol NOUN njb-49 184 8 . . PUNCT njb-49 184 9     SPACE njb-49 184 10 2 2 NUM njb-49 184 11 , , PUNCT njb-49 184 12     SPACE njb-49 184 13 no no INTJ njb-49 184 14 . . PUNCT njb-49 184 15     SPACE njb-49 185 1 1 1 NUM njb-49 185 2 : : PUNCT njb-49 185 3     SPACE njb-49 185 4 1 1 X njb-49 185 5     SPACE njb-49 185 6 – – PUNCT njb-49 185 7     SPACE njb-49 185 8 15 15 NUM njb-49 185 9                                                                                                                                                                                                                                         SPACE njb-49 185 10 biotechnology biotechnology NOUN njb-49 185 11     SPACE njb-49 185 12 society society NOUN njb-49 185 13     SPACE njb-49 185 14 of of ADP njb-49 185 15     SPACE njb-49 185 16 nepal nepal NOUN njb-49 185 17     SPACE njb-49 185 18 ( ( PUNCT njb-49 185 19 bsn bsn PROPN njb-49 185 20 ) ) PUNCT njb-49 185 21 , , PUNCT njb-49 185 22     SPACE njb-49 185 23 all all DET njb-49 185 24     SPACE njb-49 185 25 rights right NOUN njb-49 185 26     SPACE njb-49 185 27 reserved reserve VERB njb-49 185 28     SPACE njb-49 185 29 12 12 NUM njb-49 185 30 polyclonal polyclonal ADJ njb-49 185 31 antibodies antibody NOUN njb-49 185 32 exhibited exhibit VERB njb-49 185 33 a a DET njb-49 185 34 strong strong ADJ njb-49 185 35 affinity affinity NOUN njb-49 185 36 towards towards ADP njb-49 185 37 gst gst NOUN njb-49 185 38 free free ADJ njb-49 185 39 and and CCONJ njb-49 185 40 gst gst NOUN njb-49 185 41 tagged tag VERB njb-49 185 42 flc flc PROPN njb-49 185 43 and and CCONJ njb-49 185 44 f f PROPN njb-49 185 45 - - PUNCT njb-49 185 46 hn hn PROPN njb-49 185 47 respectively respectively ADV njb-49 185 48 , , PUNCT njb-49 185 49 while while SCONJ njb-49 185 50 both both PRON njb-49 185 51 recognized recognize VERB njb-49 185 52 the the DET njb-49 185 53 native native ADJ njb-49 185 54 type type NOUN njb-49 185 55 f f PROPN njb-49 185 56 toxin toxin PROPN njb-49 185 57 . . PUNCT njb-49 186 1 from from ADP njb-49 186 2 this this DET njb-49 186 3 work work NOUN njb-49 186 4 , , PUNCT njb-49 186 5 we we PRON njb-49 186 6 conclude conclude VERB njb-49 186 7 that that SCONJ njb-49 186 8 purified purify VERB njb-49 186 9 bont bont ADJ njb-49 186 10 / / SYM njb-49 186 11 f f PROPN njb-49 186 12 lc lc PROPN njb-49 186 13 and and CCONJ njb-49 186 14 hn hn PROPN njb-49 186 15 domains domain NOUN njb-49 186 16 are be AUX njb-49 186 17 capable capable ADJ njb-49 186 18 of of ADP njb-49 186 19 producing produce VERB njb-49 186 20 highly highly ADV njb-49 186 21 effective effective ADJ njb-49 186 22 immunogens immunogen NOUN njb-49 186 23 . . PUNCT njb-49 187 1 moreover moreover ADV njb-49 187 2 , , PUNCT njb-49 187 3 if if SCONJ njb-49 187 4 future future ADJ njb-49 187 5 attempts attempt NOUN njb-49 187 6 prove prove VERB njb-49 187 7 fruitful fruitful ADJ njb-49 187 8 in in ADP njb-49 187 9 the the DET njb-49 187 10 isolation isolation NOUN njb-49 187 11 of of ADP njb-49 187 12 monoclonal monoclonal NOUN njb-49 187 13 antibodies antibody NOUN njb-49 187 14 ( ( PUNCT njb-49 187 15 that that PRON njb-49 187 16 possess possess VERB njb-49 187 17 neutralizing neutralize VERB njb-49 187 18 activity activity NOUN njb-49 187 19 ) ) PUNCT njb-49 187 20 , , PUNCT njb-49 187 21 towards towards ADP njb-49 187 22 bont bont PROPN njb-49 187 23 / / SYM njb-49 187 24 f f PROPN njb-49 187 25 fragments fragment NOUN njb-49 187 26 , , PUNCT njb-49 187 27 this this PRON njb-49 187 28 may may AUX njb-49 187 29 lead lead VERB njb-49 187 30 to to ADP njb-49 187 31 more more ADV njb-49 187 32 efficient efficient ADJ njb-49 187 33 protection protection NOUN njb-49 187 34 against against ADP njb-49 187 35 a a DET njb-49 187 36 high high ADJ njb-49 187 37 neurotoxin neurotoxin NOUN njb-49 187 38 dose dose NOUN njb-49 187 39 . . PUNCT njb-49 188 1 implications implication NOUN njb-49 188 2 of of ADP njb-49 188 3 this this DET njb-49 188 4 research research NOUN njb-49 188 5 could could AUX njb-49 188 6 span span VERB njb-49 188 7 across across ADP njb-49 188 8 the the DET njb-49 188 9 development development NOUN njb-49 188 10 of of ADP njb-49 188 11 therapeutic therapeutic ADJ njb-49 188 12 approaches approach NOUN njb-49 188 13 , , PUNCT njb-49 188 14 diagnostic diagnostic ADJ njb-49 188 15 detection detection NOUN njb-49 188 16 systems system NOUN njb-49 188 17 , , PUNCT njb-49 188 18 and and CCONJ njb-49 188 19 vaccine vaccine NOUN njb-49 188 20 candidacy candidacy NOUN njb-49 188 21 for for ADP njb-49 188 22 the the DET njb-49 188 23 protection protection NOUN njb-49 188 24 and and CCONJ njb-49 188 25 treatment treatment NOUN njb-49 188 26 of of ADP njb-49 188 27 botulism botulism NOUN njb-49 188 28 as as SCONJ njb-49 188 29 has have AUX njb-49 188 30 been be AUX njb-49 188 31 elucidated elucidate VERB njb-49 188 32 in in ADP njb-49 188 33 previous previous ADJ njb-49 188 34 work work NOUN njb-49 188 35 [ [ PUNCT njb-49 188 36 34 34 NUM njb-49 188 37 , , PUNCT njb-49 188 38 37 37 NUM njb-49 188 39 , , PUNCT njb-49 188 40 39 39 NUM njb-49 188 41 - - SYM njb-49 188 42 42 42 NUM njb-49 188 43 ] ] PUNCT njb-49 188 44 . . PUNCT njb-49 189 1 in in ADP njb-49 189 2 summary summary NOUN njb-49 189 3 , , PUNCT njb-49 189 4 we we PRON njb-49 189 5 have have AUX njb-49 189 6 expressed express VERB njb-49 189 7 both both PRON njb-49 189 8 recombinant recombinant ADJ njb-49 189 9 lc lc NOUN njb-49 189 10 and and CCONJ njb-49 189 11 hn hn NOUN njb-49 189 12 domains domain NOUN njb-49 189 13 of of ADP njb-49 189 14 bont bont PROPN njb-49 189 15 / / SYM njb-49 189 16 f. f. PROPN njb-49 189 17 the the DET njb-49 189 18 recombinant recombinant ADJ njb-49 189 19 proteins protein NOUN njb-49 189 20 are be AUX njb-49 189 21 soluble soluble ADJ njb-49 189 22 ; ; PUNCT njb-49 189 23 elicit elicit VERB njb-49 189 24 an an DET njb-49 189 25 active active ADJ njb-49 189 26 immune immune ADJ njb-49 189 27 response response NOUN njb-49 189 28 ( ( PUNCT njb-49 189 29 polyclonal polyclonal ADJ njb-49 189 30 antibodies antibody NOUN njb-49 189 31 ) ) PUNCT njb-49 189 32 in in ADP njb-49 189 33 mice mouse NOUN njb-49 189 34 , , PUNCT njb-49 189 35 making make VERB njb-49 189 36 them they PRON njb-49 189 37 ideal ideal ADJ njb-49 189 38 for for SCONJ njb-49 189 39 investigators investigator NOUN njb-49 189 40 to to PART njb-49 189 41 process process VERB njb-49 189 42 potential potential ADJ njb-49 189 43 subunit subunit NOUN njb-49 189 44 vaccines vaccine NOUN njb-49 189 45 towards towards ADP njb-49 189 46 bont bont PROPN njb-49 189 47 / / SYM njb-49 189 48 f. f. PROPN njb-49 189 49 acknowledgements acknowledgement VERB njb-49 189 50 all all DET njb-49 189 51 authors author NOUN njb-49 189 52 wish wish VERB njb-49 189 53 to to PART njb-49 189 54 acknowledge acknowledge VERB njb-49 189 55 the the DET njb-49 189 56 financial financial ADJ njb-49 189 57 support support NOUN njb-49 189 58 from from ADP njb-49 189 59 medy medy PROPN njb-49 189 60 - - PUNCT njb-49 189 61 tox tox PROPN njb-49 189 62 inc inc PROPN njb-49 189 63 . . PROPN njb-49 189 64 and and CCONJ njb-49 189 65 for for ADP njb-49 189 66 their their PRON njb-49 189 67 help help NOUN njb-49 189 68 in in ADP njb-49 189 69 the the DET njb-49 189 70 acquisition acquisition NOUN njb-49 189 71 of of ADP njb-49 189 72 c. c. PROPN njb-49 189 73 botulinum botulinum PROPN njb-49 189 74 f f PROPN njb-49 189 75 str str PROPN njb-49 189 76 . . PROPN njb-49 189 77 langeland langeland PROPN njb-49 189 78 and and CCONJ njb-49 189 79 bont bont PROPN njb-49 189 80 / / SYM njb-49 189 81 f f PROPN njb-49 189 82 ( ( PUNCT njb-49 189 83 wako wako ADJ njb-49 189 84 pure pure ADJ njb-49 189 85 chemicals chemical NOUN njb-49 189 86 ind ind NOUN njb-49 189 87 . . PROPN njb-49 189 88 , , PUNCT njb-49 189 89 osaka osaka PROPN njb-49 189 90 , , PUNCT njb-49 189 91 japan japan PROPN njb-49 189 92 ) ) PUNCT njb-49 189 93 . . PUNCT njb-49 190 1 we we PRON njb-49 190 2 would would AUX njb-49 190 3 also also ADV njb-49 190 4 like like VERB njb-49 190 5 to to PART njb-49 190 6 express express VERB njb-49 190 7 our our PRON njb-49 190 8 gratitude gratitude NOUN njb-49 190 9 to to ADP njb-49 190 10 jesus jesus PROPN njb-49 190 11 andres andres PROPN njb-49 190 12 cuaron cuaron PROPN njb-49 190 13 for for ADP njb-49 190 14 his his PRON njb-49 190 15 contributions contribution NOUN njb-49 190 16 on on ADP njb-49 190 17 the the DET njb-49 190 18 drafting drafting NOUN njb-49 190 19 of of ADP njb-49 190 20 this this DET njb-49 190 21 article article NOUN njb-49 190 22 . . PUNCT njb-49 191 1 references reference NOUN njb-49 191 2 1 1 NUM njb-49 191 3 . . PUNCT njb-49 192 1 mccroskey mccroskey PROPN njb-49 192 2 lm lm PROPN njb-49 192 3 , , PUNCT njb-49 192 4 hatheway hatheway PROPN njb-49 192 5 cl cl PROPN njb-49 192 6 , , PUNCT njb-49 192 7 fenicia fenicia PROPN njb-49 192 8 l l PROPN njb-49 192 9 , , PUNCT njb-49 192 10 pasolini pasolini PROPN njb-49 192 11 b b PROPN njb-49 192 12 , , PUNCT njb-49 192 13 aureli aureli PROPN njb-49 192 14 p p NOUN njb-49 192 15 : : PUNCT njb-49 192 16 characterization characterization NOUN njb-49 192 17 of of ADP njb-49 192 18 an an DET njb-49 192 19 organism organism NOUN njb-49 192 20 that that PRON njb-49 192 21 produces produce VERB njb-49 192 22 type type NOUN njb-49 192 23 e e NOUN njb-49 192 24 botulinal botulinal ADJ njb-49 192 25 toxin toxin NOUN njb-49 192 26 but but CCONJ njb-49 192 27 which which PRON njb-49 192 28 resembles resemble VERB njb-49 192 29 c. c. PROPN njb-49 192 30 butyricum butyricum PROPN njb-49 192 31 from from ADP njb-49 192 32 the the DET njb-49 192 33 feces fece NOUN njb-49 192 34 of of ADP njb-49 192 35 an an DET njb-49 192 36 infant infant NOUN njb-49 192 37 with with ADP njb-49 192 38 type type NOUN njb-49 192 39 e e NOUN njb-49 192 40 botulism botulism NOUN njb-49 192 41 . . PUNCT njb-49 193 1 j j PROPN njb-49 193 2 clin clin PROPN njb-49 193 3 microbiol microbiol PROPN njb-49 193 4 1986 1986 NUM njb-49 193 5 , , PUNCT njb-49 193 6 23(1):201 23(1):201 PROPN njb-49 193 7 - - PUNCT njb-49 193 8 202 202 NUM njb-49 193 9 . . NOUN njb-49 194 1 2 2 X njb-49 194 2 . . PUNCT njb-49 194 3 mccroskey mccroskey PROPN njb-49 194 4 lm lm PROPN njb-49 194 5 , , PUNCT njb-49 194 6 hatheway hatheway PROPN njb-49 194 7 cl cl PROPN njb-49 194 8 , , PUNCT njb-49 194 9 woodruff woodruff PROPN njb-49 194 10 ba ba PROPN njb-49 194 11 , , PUNCT njb-49 194 12 greenberg greenberg PROPN njb-49 194 13 ja ja PROPN njb-49 194 14 , , PUNCT njb-49 194 15 jurgenson jurgenson PROPN njb-49 194 16 p p PROPN njb-49 194 17 : : PUNCT njb-49 194 18 type type PROPN njb-49 194 19 f f PROPN njb-49 194 20 botulism botulism NOUN njb-49 194 21 due due ADP njb-49 194 22 to to ADP njb-49 194 23 neurotoxigenic neurotoxigenic PROPN njb-49 194 24 c. c. PROPN njb-49 194 25 baratii baratii PROPN njb-49 194 26 from from ADP njb-49 194 27 an an DET njb-49 194 28 unknown unknown ADJ njb-49 194 29 source source NOUN njb-49 194 30 in in ADP njb-49 194 31 an an DET njb-49 194 32 adult adult NOUN njb-49 194 33 . . PUNCT njb-49 195 1 j j PROPN njb-49 195 2 clin clin PROPN njb-49 195 3 microbiol microbiol PROPN njb-49 195 4 1991 1991 NUM njb-49 195 5 , , PUNCT njb-49 195 6 29(11):26182620 29(11):26182620 NUM njb-49 195 7 . . PUNCT njb-49 195 8 3 3 X njb-49 195 9 . . PUNCT njb-49 196 1 sakaguchi sakaguchi PROPN njb-49 196 2 g g PROPN njb-49 196 3 : : PUNCT njb-49 196 4 c. c. PROPN njb-49 196 5 botulinum botulinum PROPN njb-49 196 6 toxins toxin NOUN njb-49 196 7 . . PUNCT njb-49 197 1 pharmacol pharmacol PROPN njb-49 197 2 ther ther PROPN njb-49 197 3 1982 1982 NUM njb-49 197 4 , , PUNCT njb-49 197 5 19(2):165194 19(2):165194 NUM njb-49 197 6 . . PUNCT njb-49 197 7 4 4 NUM njb-49 197 8 . . PROPN njb-49 197 9 simpson simpson PROPN njb-49 197 10 ll ll PROPN njb-49 197 11 ( ( PUNCT njb-49 197 12 ed ed NOUN njb-49 197 13 ): ): PUNCT njb-49 197 14 botulinum botulinum ADJ njb-49 197 15 neurotoxin neurotoxin NOUN njb-49 197 16 and and CCONJ njb-49 197 17 tetanus tetanus NOUN njb-49 197 18 toxin toxin NOUN njb-49 197 19 . . PUNCT njb-49 198 1 san san PROPN njb-49 198 2 diego diego PROPN njb-49 198 3 , , PUNCT njb-49 198 4 ca can AUX njb-49 198 5 : : PUNCT njb-49 198 6 academic academic ADJ njb-49 198 7 press press NOUN njb-49 198 8 ; ; PUNCT njb-49 198 9 1989 1989 NUM njb-49 198 10 . . PUNCT njb-49 199 1 5 5 X njb-49 199 2 . . PUNCT njb-49 199 3 burgen burgen PROPN njb-49 199 4 as as ADP njb-49 199 5 , , PUNCT njb-49 199 6 dickens dickens PROPN njb-49 199 7 f f PROPN njb-49 199 8 , , PUNCT njb-49 199 9 zatman zatman PROPN njb-49 199 10 lj lj PROPN njb-49 199 11 : : PUNCT njb-49 199 12 the the DET njb-49 199 13 action action NOUN njb-49 199 14 of of ADP njb-49 199 15 botulinum botulinum ADJ njb-49 199 16 toxin toxin NOUN njb-49 199 17 on on ADP njb-49 199 18 the the DET njb-49 199 19 neuro neuro PROPN njb-49 199 20 - - PUNCT njb-49 199 21 muscular muscular ADJ njb-49 199 22 junction junction NOUN njb-49 199 23 . . PUNCT njb-49 200 1 j j PROPN njb-49 200 2 physiol physiol PROPN njb-49 200 3 1949 1949 NUM njb-49 200 4 , , PUNCT njb-49 200 5 109(1 109(1 NUM njb-49 200 6 - - PUNCT njb-49 200 7 2):10 2):10 NUM njb-49 200 8 - - PUNCT njb-49 200 9 24 24 NUM njb-49 200 10 . . NUM njb-49 200 11 6 6 NUM njb-49 200 12 . . PUNCT njb-49 201 1 atlas atla NOUN njb-49 201 2 rm rm PROPN njb-49 201 3 : : PUNCT njb-49 201 4 bioterrorism bioterrorism NOUN njb-49 201 5 and and CCONJ njb-49 201 6 biodefence biodefence NOUN njb-49 201 7 research research NOUN njb-49 201 8 : : PUNCT njb-49 201 9 changing change VERB njb-49 201 10 the the DET njb-49 201 11 focus focus NOUN njb-49 201 12 of of ADP njb-49 201 13 microbiology microbiology NOUN njb-49 201 14 . . PUNCT njb-49 202 1 nat nat PROPN njb-49 202 2 rev rev PROPN njb-49 202 3 microbiol microbiol PROPN njb-49 202 4 2003 2003 NUM njb-49 202 5 , , PUNCT njb-49 202 6 1(1):70 1(1):70 NUM njb-49 202 7 - - SYM njb-49 202 8 74 74 NUM njb-49 202 9 . . NUM njb-49 202 10 7 7 NUM njb-49 202 11 . . X njb-49 202 12 bossi bossi PROPN njb-49 202 13 p p PROPN njb-49 202 14 , , PUNCT njb-49 202 15 bricaire bricaire PROPN njb-49 202 16 f f X njb-49 202 17 : : PUNCT njb-49 202 18 [ [ X njb-49 202 19 botulism botulism NOUN njb-49 202 20 toxin toxin NOUN njb-49 202 21 , , PUNCT njb-49 202 22 bioterrorist bioterrorist ADJ njb-49 202 23 weapon weapon NOUN njb-49 202 24 ] ] X njb-49 202 25 . . PUNCT njb-49 203 1 presse presse PROPN njb-49 203 2 med me VERB njb-49 203 3 2003 2003 NUM njb-49 203 4 , , PUNCT njb-49 203 5 32(10):463 32(10):463 NUM njb-49 203 6 - - SYM njb-49 203 7 465 465 NUM njb-49 203 8 . . PUNCT njb-49 204 1 nepal nepal ADJ njb-49 204 2     SPACE njb-49 204 3 journal journal NOUN njb-49 204 4     SPACE njb-49 204 5 of of ADP njb-49 204 6     SPACE njb-49 204 7 biotechnology biotechnology NOUN njb-49 204 8 . . PUNCT njb-49 204 9         SPACE njb-49 205 1 jan jan PROPN njb-49 205 2 . . PROPN njb-49 205 3     SPACE njb-49 205 4 2012 2012 NUM njb-49 205 5 , , PUNCT njb-49 205 6     SPACE njb-49 205 7 vol vol NOUN njb-49 205 8 . . PUNCT njb-49 205 9     SPACE njb-49 205 10 2 2 NUM njb-49 205 11 , , PUNCT njb-49 205 12     SPACE njb-49 205 13 no no INTJ njb-49 205 14 . . PUNCT njb-49 205 15     SPACE njb-49 206 1 1 1 NUM njb-49 206 2 : : PUNCT njb-49 206 3     SPACE njb-49 206 4 1 1 X njb-49 206 5     SPACE njb-49 206 6 – – PUNCT njb-49 206 7     SPACE njb-49 206 8 15 15 NUM njb-49 206 9                                                                                                                                                                                                                                         SPACE njb-49 206 10 biotechnology biotechnology NOUN njb-49 206 11     SPACE njb-49 206 12 society society NOUN njb-49 206 13     SPACE njb-49 206 14 of of ADP njb-49 206 15     SPACE njb-49 206 16 nepal nepal NOUN njb-49 206 17     SPACE njb-49 206 18 ( ( PUNCT njb-49 206 19 bsn bsn PROPN njb-49 206 20 ) ) PUNCT njb-49 206 21 , , PUNCT njb-49 206 22     SPACE njb-49 206 23 all all DET njb-49 206 24     SPACE njb-49 206 25 rights right NOUN njb-49 206 26     SPACE njb-49 206 27 reserved reserve VERB njb-49 206 28     SPACE njb-49 206 29 13 13 NUM njb-49 206 30 8 8 NUM njb-49 206 31 . . PUNCT njb-49 207 1 moore moore PROPN njb-49 207 2 a a PRON njb-49 207 3 : : PUNCT njb-49 207 4 the the DET njb-49 207 5 biochemistry biochemistry NOUN njb-49 207 6 of of ADP njb-49 207 7 beauty beauty NOUN njb-49 207 8 . . PUNCT njb-49 208 1 the the DET njb-49 208 2 science science NOUN njb-49 208 3 and and CCONJ njb-49 208 4 pseudoscience pseudoscience NOUN njb-49 208 5 of of ADP njb-49 208 6 beautiful beautiful ADJ njb-49 208 7 skin skin NOUN njb-49 208 8 . . PUNCT njb-49 209 1 embo embo PROPN njb-49 209 2 rep rep PROPN njb-49 209 3 2002 2002 PROPN njb-49 209 4 , , PUNCT njb-49 209 5 3(8):714 3(8):714 NUM njb-49 209 6 - - SYM njb-49 209 7 717 717 NUM njb-49 209 8 . . PUNCT njb-49 210 1 9 9 X njb-49 210 2 . . X njb-49 210 3 brin brin VERB njb-49 210 4 mf mf NOUN njb-49 210 5 : : PUNCT njb-49 210 6 botulinum botulinum ADJ njb-49 210 7 toxin toxin NOUN njb-49 210 8 : : PUNCT njb-49 210 9 chemistry chemistry NOUN njb-49 210 10 , , PUNCT njb-49 210 11 pharmacology pharmacology NOUN njb-49 210 12 , , PUNCT njb-49 210 13 toxicity toxicity NOUN njb-49 210 14 , , PUNCT njb-49 210 15 and and CCONJ njb-49 210 16 immunology immunology NOUN njb-49 210 17 . . PUNCT njb-49 211 1 muscle muscle NOUN njb-49 211 2 nerve nerve NOUN njb-49 211 3 suppl suppl NOUN njb-49 211 4 1997 1997 NUM njb-49 211 5 , , PUNCT njb-49 211 6 6 6 NUM njb-49 211 7 : : PUNCT njb-49 211 8 s146 s146 PROPN njb-49 211 9 - - PUNCT njb-49 211 10 168 168 NUM njb-49 211 11 . . NUM njb-49 211 12 10 10 NUM njb-49 211 13 . . PUNCT njb-49 212 1 dressler dressler PROPN njb-49 212 2 d d PROPN njb-49 212 3 : : PUNCT njb-49 212 4 electromyographic electromyographic ADJ njb-49 212 5 evaluation evaluation NOUN njb-49 212 6 of of ADP njb-49 212 7 cervical cervical ADJ njb-49 212 8 dystonia dystonia NOUN njb-49 212 9 for for ADP njb-49 212 10 planning planning NOUN njb-49 212 11 of of ADP njb-49 212 12 botulinum botulinum ADJ njb-49 212 13 toxin toxin NOUN njb-49 212 14 therapy therapy NOUN njb-49 212 15 . . PUNCT njb-49 213 1 eur eur PROPN njb-49 213 2 j j PROPN njb-49 213 3 neurol neurol PROPN njb-49 213 4 2000 2000 NUM njb-49 213 5 , , PUNCT njb-49 213 6 7(6):713 7(6):713 NUM njb-49 213 7 - - SYM njb-49 213 8 718 718 NUM njb-49 213 9 . . NOUN njb-49 213 10 11 11 NUM njb-49 213 11 . . PUNCT njb-49 214 1 jankovic jankovic PROPN njb-49 214 2 j j PROPN njb-49 214 3 : : PUNCT njb-49 214 4 botulinum botulinum ADJ njb-49 214 5 toxin toxin NOUN njb-49 214 6 in in ADP njb-49 214 7 movement movement NOUN njb-49 214 8 disorders disorder NOUN njb-49 214 9 . . PUNCT njb-49 215 1 curr curr PROPN njb-49 215 2 opin opin PROPN njb-49 215 3 neurol neurol PROPN njb-49 215 4 1994 1994 NUM njb-49 215 5 , , PUNCT njb-49 215 6 7(4):358 7(4):358 NUM njb-49 215 7 - - SYM njb-49 215 8 366 366 NUM njb-49 215 9 . . NUM njb-49 215 10 12 12 NUM njb-49 215 11 . . PUNCT njb-49 216 1 sugiyama sugiyama PROPN njb-49 216 2 h h PROPN njb-49 216 3 , , PUNCT njb-49 216 4 das das PROPN njb-49 216 5 gupta gupta PROPN njb-49 216 6 r r PROPN njb-49 216 7 , , PUNCT njb-49 216 8 yang yang PROPN njb-49 216 9 kh kh PROPN njb-49 216 10 : : PUNCT njb-49 216 11 disulfide disulfide NOUN njb-49 216 12 - - PUNCT njb-49 216 13 toxicity toxicity NOUN njb-49 216 14 relationship relationship NOUN njb-49 216 15 of of ADP njb-49 216 16 botulinal botulinal ADJ njb-49 216 17 toxin toxin NOUN njb-49 216 18 types type NOUN njb-49 216 19 a a DET njb-49 216 20 , , PUNCT njb-49 216 21 e e NOUN njb-49 216 22 , , PUNCT njb-49 216 23 and and CCONJ njb-49 216 24 f. f. PROPN njb-49 216 25 proc proc PROPN njb-49 216 26 soc soc PROPN njb-49 216 27 exp exp PROPN njb-49 216 28 biol biol PROPN njb-49 216 29 med me VERB njb-49 216 30 1973 1973 NUM njb-49 216 31 , , PUNCT njb-49 216 32 143(3):589 143(3):589 NUM njb-49 216 33 - - PUNCT njb-49 216 34 591 591 NUM njb-49 216 35 . . PROPN njb-49 216 36 13 13 NUM njb-49 216 37 . . PUNCT njb-49 216 38 dasgupta dasgupta PROPN njb-49 216 39 br br PROPN njb-49 216 40 , , PUNCT njb-49 216 41 sugiyama sugiyama PROPN njb-49 216 42 h h PROPN njb-49 216 43 : : PUNCT njb-49 216 44 a a DET njb-49 216 45 common common ADJ njb-49 216 46 subunit subunit NOUN njb-49 216 47 structure structure NOUN njb-49 216 48 in in ADP njb-49 216 49 c. c. PROPN njb-49 216 50 botulinum botulinum PROPN njb-49 216 51 type type VERB njb-49 216 52 a a PRON njb-49 216 53 , , PUNCT njb-49 216 54 b b NOUN njb-49 216 55 and and CCONJ njb-49 216 56 e e NOUN njb-49 216 57 toxins toxin NOUN njb-49 216 58 . . PUNCT njb-49 217 1 biochem biochem PROPN njb-49 217 2 biophys biophys PROPN njb-49 217 3 res res PROPN njb-49 217 4 commun commun PROPN njb-49 217 5 1972 1972 NUM njb-49 217 6 , , PUNCT njb-49 217 7 48(1):108 48(1):108 NUM njb-49 217 8 - - SYM njb-49 217 9 112 112 NUM njb-49 217 10 . . NUM njb-49 217 11 14 14 NUM njb-49 217 12 . . PUNCT njb-49 218 1 schmidt schmidt PROPN njb-49 218 2 jj jj PROPN njb-49 218 3 , , PUNCT njb-49 218 4 stafford stafford PROPN njb-49 218 5 rg rg PROPN njb-49 218 6 : : PUNCT njb-49 218 7 botulinum botulinum PROPN njb-49 218 8 neurotoxin neurotoxin NOUN njb-49 218 9 serotype serotype VERB njb-49 218 10 f f PROPN njb-49 218 11 : : PUNCT njb-49 218 12 identification identification NOUN njb-49 218 13 of of ADP njb-49 218 14 substrate substrate ADJ njb-49 218 15 recognition recognition NOUN njb-49 218 16 requirements requirement NOUN njb-49 218 17 and and CCONJ njb-49 218 18 development development NOUN njb-49 218 19 of of ADP njb-49 218 20 inhibitors inhibitor NOUN njb-49 218 21 with with ADP njb-49 218 22 low low ADJ njb-49 218 23 nanomolar nanomolar ADJ njb-49 218 24 affinity affinity NOUN njb-49 218 25 . . PUNCT njb-49 219 1 biochemistry biochemistry NOUN njb-49 219 2 2005 2005 NUM njb-49 219 3 , , PUNCT njb-49 219 4 44(10):4067 44(10):4067 NUM njb-49 219 5 - - SYM njb-49 219 6 4073 4073 NUM njb-49 219 7 . . PUNCT njb-49 220 1 15 15 X njb-49 220 2 . . X njb-49 220 3 schiavo schiavo PROPN njb-49 220 4 g g PROPN njb-49 220 5 , , PUNCT njb-49 220 6 rossetto rossetto PROPN njb-49 220 7 o o PROPN njb-49 220 8 , , PUNCT njb-49 220 9 catsicas catsicas PROPN njb-49 220 10 s s PROPN njb-49 220 11 , , PUNCT njb-49 220 12 polverino polverino PROPN njb-49 220 13 de de X njb-49 220 14 laureto laureto PROPN njb-49 220 15 p p PROPN njb-49 220 16 , , PUNCT njb-49 220 17 dasgupta dasgupta PROPN njb-49 220 18 br br PROPN njb-49 220 19 , , PUNCT njb-49 220 20 benfenati benfenati PROPN njb-49 220 21 f f PROPN njb-49 220 22 , , PUNCT njb-49 220 23 montecucco montecucco PROPN njb-49 220 24 c c PROPN njb-49 220 25 : : PUNCT njb-49 220 26 identification identification NOUN njb-49 220 27 of of ADP njb-49 220 28 the the DET njb-49 220 29 nerve nerve NOUN njb-49 220 30 terminal terminal NOUN njb-49 220 31 targets target NOUN njb-49 220 32 of of ADP njb-49 220 33 botulinum botulinum ADJ njb-49 220 34 neurotoxin neurotoxin NOUN njb-49 220 35 serotypes serotype VERB njb-49 220 36 a a DET njb-49 220 37 , , PUNCT njb-49 220 38 d d PROPN njb-49 220 39 , , PUNCT njb-49 220 40 and and CCONJ njb-49 220 41 e. e. PROPN njb-49 220 42 j j PROPN njb-49 220 43 biol biol PROPN njb-49 220 44 chem chem PROPN njb-49 220 45 1993 1993 NUM njb-49 220 46 , , PUNCT njb-49 220 47 268(32):23784 268(32):23784 NUM njb-49 220 48 - - SYM njb-49 220 49 23787 23787 NUM njb-49 220 50 . . PUNCT njb-49 221 1 16 16 NUM njb-49 221 2 . . PUNCT njb-49 222 1 chen chen PROPN njb-49 222 2 s s PROPN njb-49 222 3 , , PUNCT njb-49 222 4 wan wan PROPN njb-49 222 5 hy hy PROPN njb-49 222 6 : : PUNCT njb-49 222 7 molecular molecular ADJ njb-49 222 8 mechanisms mechanism NOUN njb-49 222 9 of of ADP njb-49 222 10 substrate substrate ADJ njb-49 222 11 recognition recognition NOUN njb-49 222 12 and and CCONJ njb-49 222 13 specificity specificity NOUN njb-49 222 14 of of ADP njb-49 222 15 botulinum botulinum ADJ njb-49 222 16 neurotoxin neurotoxin NOUN njb-49 222 17 serotype serotype PROPN njb-49 222 18 f. f. PROPN njb-49 222 19 biochem biochem PROPN njb-49 222 20 j j PROPN njb-49 222 21 2011 2011 NUM njb-49 222 22 , , PUNCT njb-49 222 23 433(2):277 433(2):277 NUM njb-49 222 24 - - SYM njb-49 222 25 284 284 NUM njb-49 222 26 . . NUM njb-49 222 27 17 17 NUM njb-49 222 28 . . PUNCT njb-49 223 1 agarwal agarwal PROPN njb-49 223 2 r r PROPN njb-49 223 3 , , PUNCT njb-49 223 4 schmidt schmidt PROPN njb-49 223 5 jj jj PROPN njb-49 223 6 , , PUNCT njb-49 223 7 stafford stafford PROPN njb-49 223 8 rg rg PROPN njb-49 223 9 , , PUNCT njb-49 223 10 swaminathan swaminathan PROPN njb-49 223 11 s s PROPN njb-49 223 12 : : PUNCT njb-49 223 13 mode mode NOUN njb-49 223 14 of of ADP njb-49 223 15 vamp vamp NOUN njb-49 223 16 substrate substrate NOUN njb-49 223 17 recognition recognition NOUN njb-49 223 18 and and CCONJ njb-49 223 19 inhibition inhibition NOUN njb-49 223 20 of of ADP njb-49 223 21 c. c. PROPN njb-49 223 22 botulinum botulinum PROPN njb-49 223 23 neurotoxin neurotoxin PROPN njb-49 223 24 f. f. PROPN njb-49 223 25 nat nat PROPN njb-49 223 26 struct struct PROPN njb-49 223 27 mol mol PROPN njb-49 223 28 biol biol NOUN njb-49 223 29 2009 2009 NUM njb-49 223 30 , , PUNCT njb-49 223 31 16(7):789 16(7):789 NUM njb-49 223 32 - - PUNCT njb-49 223 33 794 794 NUM njb-49 223 34 . . PROPN njb-49 223 35 18 18 NUM njb-49 223 36 . . PUNCT njb-49 224 1 montecucco montecucco PROPN njb-49 224 2 c c PROPN njb-49 224 3 , , PUNCT njb-49 224 4 schiavo schiavo PROPN njb-49 224 5 g g PROPN njb-49 224 6 : : PUNCT njb-49 224 7 mechanism mechanism NOUN njb-49 224 8 of of ADP njb-49 224 9 action action NOUN njb-49 224 10 of of ADP njb-49 224 11 tetanus tetanus NOUN njb-49 224 12 and and CCONJ njb-49 224 13 botulinum botulinum NOUN njb-49 224 14 neurotoxins neurotoxin NOUN njb-49 224 15 . . PUNCT njb-49 225 1 mol mol PROPN njb-49 225 2 microbiol microbiol PROPN njb-49 225 3 1994 1994 NUM njb-49 225 4 , , PUNCT njb-49 225 5 13(1):1 13(1):1 NUM njb-49 225 6 - - SYM njb-49 225 7 8 8 NUM njb-49 225 8 . . NUM njb-49 225 9 19 19 NUM njb-49 225 10 . . PUNCT njb-49 226 1 schiavo schiavo PROPN njb-49 226 2 g g PROPN njb-49 226 3 , , PUNCT njb-49 226 4 matteoli matteoli PROPN njb-49 226 5 m m PROPN njb-49 226 6 , , PUNCT njb-49 226 7 montecucco montecucco PROPN njb-49 226 8 c c PROPN njb-49 226 9 : : PUNCT njb-49 226 10 neurotoxins neurotoxin NOUN njb-49 226 11 affecting affect VERB njb-49 226 12 neuroexocytosis neuroexocytosis NOUN njb-49 226 13 . . PUNCT njb-49 227 1 physiol physiol PROPN njb-49 227 2 rev rev PROPN njb-49 227 3 2000 2000 NUM njb-49 227 4 , , PUNCT njb-49 227 5 80(2):717 80(2):717 NUM njb-49 227 6 - - PUNCT njb-49 227 7 766 766 NUM njb-49 227 8 . . NOUN njb-49 227 9 20 20 NUM njb-49 227 10 . . PUNCT njb-49 228 1 lacy lacy PROPN njb-49 228 2 db db PROPN njb-49 228 3 , , PUNCT njb-49 228 4 tepp tepp PROPN njb-49 228 5 w w PROPN njb-49 228 6 , , PUNCT njb-49 228 7 cohen cohen PROPN njb-49 228 8 ac ac PROPN njb-49 228 9 , , PUNCT njb-49 228 10 dasgupta dasgupta PROPN njb-49 228 11 br br PROPN njb-49 228 12 , , PUNCT njb-49 228 13 stevens stevens PROPN njb-49 228 14 rc rc PROPN njb-49 228 15 : : PUNCT njb-49 228 16 crystal crystal PROPN njb-49 228 17 structure structure NOUN njb-49 228 18 of of ADP njb-49 228 19 botulinum botulinum ADJ njb-49 228 20 neurotoxin neurotoxin NOUN njb-49 228 21 type type VERB njb-49 228 22 a a PRON njb-49 228 23 and and CCONJ njb-49 228 24 implications implication NOUN njb-49 228 25 for for ADP njb-49 228 26 toxicity toxicity NOUN njb-49 228 27 . . PUNCT njb-49 229 1 nat nat PROPN njb-49 229 2 struct struct VERB njb-49 229 3 biol biol PROPN njb-49 229 4 1998 1998 NUM njb-49 229 5 , , PUNCT njb-49 229 6 5(10):898 5(10):898 NUM njb-49 229 7 - - SYM njb-49 229 8 902 902 NUM njb-49 229 9 . . PUNCT njb-49 229 10 21 21 NUM njb-49 229 11 . . PUNCT njb-49 230 1 montecucco montecucco PROPN njb-49 230 2 c c PROPN njb-49 230 3 , , PUNCT njb-49 230 4 schiavo schiavo PROPN njb-49 230 5 g g PROPN njb-49 230 6 , , PUNCT njb-49 230 7 gao gao PROPN njb-49 230 8 z z PROPN njb-49 230 9 , , PUNCT njb-49 230 10 bauerlein bauerlein PROPN njb-49 230 11 e e PROPN njb-49 230 12 , , PUNCT njb-49 230 13 boquet boquet PROPN njb-49 230 14 p p PROPN njb-49 230 15 , , PUNCT njb-49 230 16 dasgupta dasgupta PROPN njb-49 230 17 br br PROPN njb-49 230 18 : : PUNCT njb-49 230 19 interaction interaction NOUN njb-49 230 20 of of ADP njb-49 230 21 botulinum botulinum NOUN njb-49 230 22 and and CCONJ njb-49 230 23 tetanus tetanus NOUN njb-49 230 24 toxins toxin NOUN njb-49 230 25 with with ADP njb-49 230 26 the the DET njb-49 230 27 lipid lipid ADJ njb-49 230 28 bilayer bilayer NOUN njb-49 230 29 surface surface NOUN njb-49 230 30 . . PUNCT njb-49 231 1 biochem biochem PROPN njb-49 231 2 j j PROPN njb-49 231 3 1988 1988 NUM njb-49 231 4 , , PUNCT njb-49 231 5 251(2):379 251(2):379 NUM njb-49 231 6 - - SYM njb-49 231 7 383 383 NUM njb-49 231 8 . . PUNCT njb-49 232 1 22 22 NUM njb-49 232 2 . . PUNCT njb-49 233 1 yu yu PROPN njb-49 233 2 yz yz PROPN njb-49 233 3 , , PUNCT njb-49 233 4 sun sun PROPN njb-49 233 5 zw zw PROPN njb-49 233 6 , , PUNCT njb-49 233 7 wang wang PROPN njb-49 233 8 s s PROPN njb-49 233 9 , , PUNCT njb-49 233 10 yu yu PROPN njb-49 233 11 wy wy PROPN njb-49 233 12 : : PUNCT njb-49 233 13 high high ADJ njb-49 233 14 - - PUNCT njb-49 233 15 level level NOUN njb-49 233 16 expression expression NOUN njb-49 233 17 of of ADP njb-49 233 18 the the DET njb-49 233 19 hcc hcc PROPN njb-49 233 20 domain domain NOUN njb-49 233 21 of of ADP njb-49 233 22 c. c. PROPN njb-49 233 23 botulinum botulinum PROPN njb-49 233 24 neurotoxin neurotoxin NOUN njb-49 233 25 serotype serotype VERB njb-49 233 26 a a PRON njb-49 233 27 in in ADP njb-49 233 28 e. e. PROPN njb-49 233 29 coli coli PROPN njb-49 233 30 and and CCONJ njb-49 233 31 its its PRON njb-49 233 32 immunogenicity immunogenicity NOUN njb-49 233 33 as as ADP njb-49 233 34 an an DET njb-49 233 35 antigen antigen NOUN njb-49 233 36 . . PUNCT njb-49 234 1 sheng sheng PROPN njb-49 234 2 wu wu PROPN njb-49 234 3 gong gong PROPN njb-49 234 4 cheng cheng PROPN njb-49 234 5 xue xue PROPN njb-49 234 6 bao bao PROPN njb-49 234 7 2007 2007 NUM njb-49 234 8 , , PUNCT njb-49 234 9 23(5):812 23(5):812 NUM njb-49 234 10 - - PUNCT njb-49 234 11 817 817 NUM njb-49 234 12 . . PUNCT njb-49 234 13 23 23 NUM njb-49 234 14 . . PUNCT njb-49 235 1 moller moller PROPN njb-49 235 2 v v PROPN njb-49 235 3 , , PUNCT njb-49 235 4 scheibel scheibel PROPN njb-49 235 5 i i PRON njb-49 235 6 : : PUNCT njb-49 235 7 preliminary preliminary ADJ njb-49 235 8 report report NOUN njb-49 235 9 on on ADP njb-49 235 10 the the DET njb-49 235 11 isolation isolation NOUN njb-49 235 12 of of ADP njb-49 235 13 an an DET njb-49 235 14 apparently apparently ADV njb-49 235 15 new new ADJ njb-49 235 16 type type NOUN njb-49 235 17 of of ADP njb-49 235 18 c. c. PROPN njb-49 235 19 botulinum botulinum PROPN njb-49 235 20 . . PUNCT njb-49 236 1 acta acta PROPN njb-49 236 2 pathol pathol PROPN njb-49 236 3 microbiol microbiol PROPN njb-49 236 4 scand scand NOUN njb-49 236 5 1960 1960 NUM njb-49 236 6 , , PUNCT njb-49 236 7 48:80 48:80 NUM njb-49 236 8 . . PUNCT njb-49 237 1 nepal nepal ADJ njb-49 237 2     SPACE njb-49 237 3 journal journal NOUN njb-49 237 4     SPACE njb-49 237 5 of of ADP njb-49 237 6     SPACE njb-49 237 7 biotechnology biotechnology NOUN njb-49 237 8 . . PUNCT njb-49 237 9         SPACE njb-49 238 1 jan jan PROPN njb-49 238 2 . . PROPN njb-49 238 3     SPACE njb-49 238 4 2012 2012 NUM njb-49 238 5 , , PUNCT njb-49 238 6     SPACE njb-49 238 7 vol vol NOUN njb-49 238 8 . . PUNCT njb-49 238 9     SPACE njb-49 238 10 2 2 NUM njb-49 238 11 , , PUNCT njb-49 238 12     SPACE njb-49 238 13 no no INTJ njb-49 238 14 . . PUNCT njb-49 238 15     SPACE njb-49 239 1 1 1 NUM njb-49 239 2 : : PUNCT njb-49 239 3     SPACE njb-49 239 4 1 1 X njb-49 239 5     SPACE njb-49 239 6 – – PUNCT njb-49 239 7     SPACE njb-49 239 8 15 15 NUM njb-49 239 9                                                                                                                                                                                                                                         SPACE njb-49 239 10 biotechnology biotechnology NOUN njb-49 239 11     SPACE njb-49 239 12 society society NOUN njb-49 239 13     SPACE njb-49 239 14 of of ADP njb-49 239 15     SPACE njb-49 239 16 nepal nepal NOUN njb-49 239 17     SPACE njb-49 239 18 ( ( PUNCT njb-49 239 19 bsn bsn PROPN njb-49 239 20 ) ) PUNCT njb-49 239 21 , , PUNCT njb-49 239 22     SPACE njb-49 239 23 all all DET njb-49 239 24     SPACE njb-49 239 25 rights right NOUN njb-49 239 26     SPACE njb-49 239 27 reserved reserve VERB njb-49 239 28     SPACE njb-49 239 29 14 14 NUM njb-49 239 30 24 24 NUM njb-49 239 31 . . PUNCT njb-49 240 1 kohler kohler NOUN njb-49 240 2 g g PROPN njb-49 240 3 , , PUNCT njb-49 240 4 howe howe PROPN njb-49 240 5 sc sc PROPN njb-49 240 6 , , PUNCT njb-49 240 7 milstein milstein PROPN njb-49 240 8 c c PROPN njb-49 240 9 : : PUNCT njb-49 240 10 fusion fusion NOUN njb-49 240 11 between between ADP njb-49 240 12 immunoglobulinsecreting immunoglobulinsecrete VERB njb-49 240 13 and and CCONJ njb-49 240 14 nonsecreting nonsecreting ADJ njb-49 240 15 myeloma myeloma NOUN njb-49 240 16 cell cell NOUN njb-49 240 17 lines line NOUN njb-49 240 18 . . PUNCT njb-49 241 1 eur eur PROPN njb-49 241 2 j j PROPN njb-49 241 3 immunol immunol NOUN njb-49 241 4 1976 1976 NUM njb-49 241 5 , , PUNCT njb-49 241 6 6(4):292 6(4):292 NUM njb-49 241 7 - - SYM njb-49 241 8 295 295 NUM njb-49 241 9 . . PUNCT njb-49 241 10 25 25 NUM njb-49 241 11 . . PUNCT njb-49 242 1 harlow harlow PROPN njb-49 242 2 e e PROPN njb-49 242 3 , , PUNCT njb-49 242 4 lane lane PROPN njb-49 242 5 d d NOUN njb-49 242 6 : : PUNCT njb-49 242 7 antibodies antibody NOUN njb-49 242 8 : : PUNCT njb-49 242 9 a a DET njb-49 242 10 laboratory laboratory NOUN njb-49 242 11 manual manual NOUN njb-49 242 12 : : PUNCT njb-49 242 13 cold cold ADJ njb-49 242 14 spring spring NOUN njb-49 242 15 harbor harbor PROPN njb-49 242 16 laboratory laboratory PROPN njb-49 242 17 press press PROPN njb-49 242 18 ; ; PUNCT njb-49 242 19 1988 1988 NUM njb-49 242 20 . . PUNCT njb-49 243 1 26 26 NUM njb-49 243 2 . . PUNCT njb-49 243 3 towbin towbin PROPN njb-49 243 4 h h PROPN njb-49 243 5 , , PUNCT njb-49 243 6 staehelin staehelin PROPN njb-49 243 7 t t PROPN njb-49 243 8 , , PUNCT njb-49 243 9 gordon gordon PROPN njb-49 243 10 j j PROPN njb-49 243 11 : : PUNCT njb-49 243 12 electrophoretic electrophoretic ADJ njb-49 243 13 transfer transfer NOUN njb-49 243 14 of of ADP njb-49 243 15 proteins protein NOUN njb-49 243 16 from from ADP njb-49 243 17 polyacrylamide polyacrylamide NOUN njb-49 243 18 gels gel NOUN njb-49 243 19 to to ADP njb-49 243 20 nitrocellulose nitrocellulose NOUN njb-49 243 21 sheets sheet NOUN njb-49 243 22 : : PUNCT njb-49 243 23 procedure procedure NOUN njb-49 243 24 and and CCONJ njb-49 243 25 some some DET njb-49 243 26 applications application NOUN njb-49 243 27 . . PUNCT njb-49 244 1 proc proc PROPN njb-49 244 2 natl natl PROPN njb-49 244 3 acad acad PROPN njb-49 244 4 sci sci PROPN njb-49 244 5 u u PROPN njb-49 244 6 s s VERB njb-49 245 1 a a PRON njb-49 245 2 1979 1979 NUM njb-49 245 3 , , PUNCT njb-49 245 4 76(9):43504354 76(9):43504354 NUM njb-49 245 5 . . PUNCT njb-49 245 6 27 27 NUM njb-49 245 7 . . PUNCT njb-49 246 1 agarwal agarwal PROPN njb-49 246 2 r r PROPN njb-49 246 3 , , PUNCT njb-49 246 4 eswaramoorthy eswaramoorthy PROPN njb-49 246 5 s s PROPN njb-49 246 6 , , PUNCT njb-49 246 7 kumaran kumaran PROPN njb-49 246 8 d d PROPN njb-49 246 9 , , PUNCT njb-49 246 10 dunn dunn PROPN njb-49 246 11 jj jj PROPN njb-49 246 12 , , PUNCT njb-49 246 13 swaminathan swaminathan PROPN njb-49 246 14 s s PROPN njb-49 246 15 : : PUNCT njb-49 246 16 cloning cloning NOUN njb-49 246 17 , , PUNCT njb-49 246 18 high high ADJ njb-49 246 19 level level NOUN njb-49 246 20 expression expression NOUN njb-49 246 21 , , PUNCT njb-49 246 22 purification purification NOUN njb-49 246 23 , , PUNCT njb-49 246 24 and and CCONJ njb-49 246 25 crystallization crystallization NOUN njb-49 246 26 of of ADP njb-49 246 27 the the DET njb-49 246 28 full full ADJ njb-49 246 29 length length NOUN njb-49 246 30 c. c. PROPN njb-49 246 31 botulinum botulinum PROPN njb-49 246 32 neurotoxin neurotoxin NOUN njb-49 246 33 type type VERB njb-49 246 34 e e NOUN njb-49 246 35 light light NOUN njb-49 246 36 chain chain NOUN njb-49 246 37 . . PUNCT njb-49 247 1 protein protein NOUN njb-49 247 2 expr expr VERB njb-49 247 3 purif purif PROPN njb-49 247 4 2004 2004 NUM njb-49 247 5 , , PUNCT njb-49 247 6 34(1):95102 34(1):95102 NUM njb-49 247 7 . . PUNCT njb-49 248 1 28 28 NUM njb-49 248 2 . . PUNCT njb-49 249 1 zhou zhou PROPN njb-49 249 2 y y PROPN njb-49 249 3 , , PUNCT njb-49 249 4 singh singh PROPN njb-49 249 5 br br PROPN njb-49 249 6 : : PUNCT njb-49 249 7 cloning cloning NOUN njb-49 249 8 , , PUNCT njb-49 249 9 highlevel highlevel NOUN njb-49 249 10 expression expression NOUN njb-49 249 11 , , PUNCT njb-49 249 12 single single ADJ njb-49 249 13 - - PUNCT njb-49 249 14 step step NOUN njb-49 249 15 purification purification NOUN njb-49 249 16 , , PUNCT njb-49 249 17 and and CCONJ njb-49 249 18 binding bind VERB njb-49 249 19 activity activity NOUN njb-49 249 20 of of ADP njb-49 249 21 his6 his6 PROPN njb-49 249 22 - - PUNCT njb-49 249 23 tagged tag VERB njb-49 249 24 recombinant recombinant ADJ njb-49 249 25 type type NOUN njb-49 249 26 b b PROPN njb-49 249 27 botulinum botulinum NOUN njb-49 249 28 neurotoxin neurotoxin VERB njb-49 249 29 heavy heavy ADJ njb-49 249 30 chain chain NOUN njb-49 249 31 transmembrane transmembrane NOUN njb-49 249 32 and and CCONJ njb-49 249 33 binding bind VERB njb-49 249 34 domain domain NOUN njb-49 249 35 . . PUNCT njb-49 250 1 protein protein NOUN njb-49 250 2 expr expr VERB njb-49 250 3 purif purif PROPN njb-49 250 4 2004 2004 NUM njb-49 250 5 , , PUNCT njb-49 250 6 34(1):8 34(1):8 PROPN njb-49 250 7 - - SYM njb-49 250 8 16 16 NUM njb-49 250 9 . . NUM njb-49 250 10 29 29 NUM njb-49 250 11 . . PUNCT njb-49 251 1 sharma sharma PROPN njb-49 251 2 s s PROPN njb-49 251 3 , , PUNCT njb-49 251 4 zhou zhou PROPN njb-49 251 5 y y PROPN njb-49 251 6 , , PUNCT njb-49 251 7 singh singh PROPN njb-49 251 8 br br PROPN njb-49 251 9 : : PUNCT njb-49 251 10 cloning cloning NOUN njb-49 251 11 , , PUNCT njb-49 251 12 expression expression NOUN njb-49 251 13 , , PUNCT njb-49 251 14 and and CCONJ njb-49 251 15 purification purification NOUN njb-49 251 16 of of ADP njb-49 251 17 c c NOUN njb-49 251 18 - - PUNCT njb-49 251 19 terminal terminal ADJ njb-49 251 20 quarter quarter NOUN njb-49 251 21 of of ADP njb-49 251 22 the the DET njb-49 251 23 heavy heavy ADJ njb-49 251 24 chain chain NOUN njb-49 251 25 of of ADP njb-49 251 26 botulinum botulinum ADJ njb-49 251 27 neurotoxin neurotoxin NOUN njb-49 251 28 type type NOUN njb-49 251 29 a. a. NOUN njb-49 251 30 protein protein PROPN njb-49 251 31 expr expr VERB njb-49 251 32 purif purif PROPN njb-49 251 33 2006 2006 NUM njb-49 251 34 , , PUNCT njb-49 251 35 45(2):288 45(2):288 PROPN njb-49 251 36 - - PUNCT njb-49 251 37 295 295 NUM njb-49 251 38 . . PUNCT njb-49 251 39 30 30 NUM njb-49 251 40 . . PUNCT njb-49 252 1 mansour mansour PROPN njb-49 252 2 aa aa PROPN njb-49 252 3 , , PUNCT njb-49 252 4 mousavi mousavi PROPN njb-49 252 5 sl sl PROPN njb-49 252 6 , , PUNCT njb-49 252 7 rasooli rasooli PROPN njb-49 252 8 i i PROPN njb-49 252 9 , , PUNCT njb-49 252 10 nazarian nazarian PROPN njb-49 252 11 s s PROPN njb-49 252 12 , , PUNCT njb-49 252 13 amani amani PROPN njb-49 252 14 j j PROPN njb-49 252 15 , , PUNCT njb-49 252 16 farhadi farhadi PROPN njb-49 252 17 n n NOUN njb-49 252 18 : : PUNCT njb-49 252 19 cloning cloning NOUN njb-49 252 20 , , PUNCT njb-49 252 21 high high ADJ njb-49 252 22 level level NOUN njb-49 252 23 expression expression NOUN njb-49 252 24 and and CCONJ njb-49 252 25 immunogenicity immunogenicity NOUN njb-49 252 26 of of ADP njb-49 252 27 1163 1163 NUM njb-49 252 28 - - SYM njb-49 252 29 1256 1256 NUM njb-49 252 30 residues residue NOUN njb-49 252 31 of of ADP njb-49 252 32 c c ADJ njb-49 252 33 - - PUNCT njb-49 252 34 terminal terminal ADJ njb-49 252 35 heavy heavy ADJ njb-49 252 36 chain chain NOUN njb-49 252 37 of of ADP njb-49 252 38 c. c. PROPN njb-49 252 39 botulinum botulinum PROPN njb-49 252 40 neurotoxin neurotoxin NOUN njb-49 252 41 type type NOUN njb-49 252 42 e. e. PROPN njb-49 252 43 biologicals biological VERB njb-49 252 44 2010 2010 NUM njb-49 252 45 , , PUNCT njb-49 252 46 38(2):260 38(2):260 NOUN njb-49 252 47 - - SYM njb-49 252 48 264 264 NUM njb-49 252 49 . . NOUN njb-49 252 50 31 31 NUM njb-49 252 51 . . PUNCT njb-49 253 1 whelan whelan PROPN njb-49 253 2 sm sm PROPN njb-49 253 3 , , PUNCT njb-49 253 4 elmore elmore PROPN njb-49 253 5 mj mj PROPN njb-49 253 6 , , PUNCT njb-49 253 7 bodsworth bodsworth PROPN njb-49 253 8 nj nj PROPN njb-49 253 9 , , PUNCT njb-49 253 10 brehm brehm PROPN njb-49 253 11 jk jk PROPN njb-49 253 12 , , PUNCT njb-49 253 13 atkinson atkinson PROPN njb-49 253 14 t t PROPN njb-49 253 15 , , PUNCT njb-49 253 16 minton minton PROPN njb-49 253 17 np np PROPN njb-49 253 18 : : PUNCT njb-49 253 19 molecular molecular ADJ njb-49 253 20 cloning cloning NOUN njb-49 253 21 of of ADP njb-49 253 22 the the DET njb-49 253 23 c. c. PROPN njb-49 253 24 botulinum botulinum PROPN njb-49 253 25 structural structural ADJ njb-49 253 26 gene gene NOUN njb-49 253 27 encoding encode VERB njb-49 253 28 the the DET njb-49 253 29 type type NOUN njb-49 253 30 b b PROPN njb-49 253 31 neurotoxin neurotoxin NOUN njb-49 253 32 and and CCONJ njb-49 253 33 determination determination NOUN njb-49 253 34 of of ADP njb-49 253 35 its its PRON njb-49 253 36 entire entire ADJ njb-49 253 37 nucleotide nucleotide NOUN njb-49 253 38 sequence sequence NOUN njb-49 253 39 . . PUNCT njb-49 254 1 appl appl VERB njb-49 254 2 environ environ NOUN njb-49 254 3 microbiol microbiol ADJ njb-49 254 4 1992 1992 NUM njb-49 254 5 , , PUNCT njb-49 254 6 58(8):2345 58(8):2345 NUM njb-49 254 7 - - PUNCT njb-49 254 8 2354 2354 NUM njb-49 254 9 . . PUNCT njb-49 255 1 32 32 NUM njb-49 255 2 . . PUNCT njb-49 256 1 yang yang PROPN njb-49 256 2 gh gh PROPN njb-49 256 3 , , PUNCT njb-49 256 4 rhee rhee PROPN njb-49 256 5 sd sd PROPN njb-49 256 6 , , PUNCT njb-49 256 7 jung jung PROPN njb-49 256 8 hh hh INTJ njb-49 256 9 , , PUNCT njb-49 256 10 jhee jhee PROPN njb-49 257 1 oh oh INTJ njb-49 257 2 , , PUNCT njb-49 257 3 yang yang PROPN njb-49 257 4 kh kh PROPN njb-49 257 5 : : PUNCT njb-49 257 6 cloning cloning NOUN njb-49 257 7 and and CCONJ njb-49 257 8 characterization characterization NOUN njb-49 257 9 of of ADP njb-49 257 10 the the DET njb-49 257 11 upstream upstream ADJ njb-49 257 12 region region NOUN njb-49 257 13 of of ADP njb-49 257 14 c. c. PROPN njb-49 257 15 botulinum botulinum PROPN njb-49 257 16 type type PROPN njb-49 257 17 b b PROPN njb-49 257 18 neurotoxin neurotoxin NOUN njb-49 257 19 gene gene NOUN njb-49 257 20 . . PUNCT njb-49 258 1 biochem biochem PROPN njb-49 258 2 mol mol PROPN njb-49 258 3 biol biol VERB njb-49 258 4 int int PROPN njb-49 258 5 1998 1998 NUM njb-49 258 6 , , PUNCT njb-49 258 7 45(2):401 45(2):401 NOUN njb-49 258 8 - - PUNCT njb-49 258 9 407 407 NUM njb-49 258 10 . . PUNCT njb-49 258 11 33 33 NUM njb-49 258 12 . . PUNCT njb-49 259 1 zhou zhou PROPN njb-49 259 2 y y PROPN njb-49 259 3 , , PUNCT njb-49 259 4 paturi paturi PROPN njb-49 259 5 s s PROPN njb-49 259 6 , , PUNCT njb-49 259 7 lindo lindo PROPN njb-49 259 8 p p PROPN njb-49 259 9 , , PUNCT njb-49 259 10 shoesmith shoesmith PROPN njb-49 259 11 sm sm PROPN njb-49 259 12 , , PUNCT njb-49 259 13 singh singh PROPN njb-49 259 14 br br PROPN njb-49 259 15 : : PUNCT njb-49 259 16 cloning cloning NOUN njb-49 259 17 , , PUNCT njb-49 259 18 expression expression NOUN njb-49 259 19 , , PUNCT njb-49 259 20 purification purification NOUN njb-49 259 21 , , PUNCT njb-49 259 22 and and CCONJ njb-49 259 23 characterization characterization NOUN njb-49 259 24 of of ADP njb-49 259 25 biologically biologically ADV njb-49 259 26 active active ADJ njb-49 259 27 recombinant recombinant ADJ njb-49 259 28 hemagglutinin33 hemagglutinin33 NOUN njb-49 259 29 , , PUNCT njb-49 259 30 type type VERB njb-49 259 31 a a DET njb-49 259 32 botulinum botulinum ADJ njb-49 259 33 neurotoxin neurotoxin NOUN njb-49 259 34 associated associate VERB njb-49 259 35 protein protein NOUN njb-49 259 36 . . PUNCT njb-49 260 1 protein protein PROPN njb-49 260 2 j j PROPN njb-49 260 3 2007 2007 NUM njb-49 260 4 , , PUNCT njb-49 260 5 26(1):29 26(1):29 NUM njb-49 260 6 - - SYM njb-49 260 7 37 37 NUM njb-49 260 8 . . PUNCT njb-49 260 9 34 34 NUM njb-49 260 10 . . PUNCT njb-49 261 1 yu yu PROPN njb-49 261 2 yz yz PROPN njb-49 261 3 , , PUNCT njb-49 261 4 li li PROPN njb-49 261 5 n n PROPN njb-49 261 6 , , PUNCT njb-49 261 7 wang wang PROPN njb-49 261 8 rl rl PROPN njb-49 261 9 , , PUNCT njb-49 261 10 zhu zhu PROPN njb-49 261 11 hq hq PROPN njb-49 261 12 , , PUNCT njb-49 261 13 wang wang PROPN njb-49 261 14 s s PROPN njb-49 261 15 , , PUNCT njb-49 261 16 yu yu PROPN njb-49 261 17 wy wy PROPN njb-49 261 18 , , PUNCT njb-49 261 19 sun sun PROPN njb-49 261 20 zw zw PROPN njb-49 261 21 : : PUNCT njb-49 261 22 evaluation evaluation NOUN njb-49 261 23 of of ADP njb-49 261 24 a a DET njb-49 261 25 recombinant recombinant ADJ njb-49 261 26 hc hc NOUN njb-49 261 27 of of ADP njb-49 261 28 c. c. PROPN njb-49 261 29 botulinum botulinum PROPN njb-49 261 30 neurotoxin neurotoxin NOUN njb-49 261 31 serotype serotype VERB njb-49 261 32 f f PROPN njb-49 261 33 as as ADP njb-49 261 34 an an DET njb-49 261 35 effective effective ADJ njb-49 261 36 subunit subunit NOUN njb-49 261 37 vaccine vaccine NOUN njb-49 261 38 . . PUNCT njb-49 262 1 clin clin PROPN njb-49 262 2 vaccine vaccine PROPN njb-49 262 3 immunol immunol NOUN njb-49 262 4 2008 2008 NUM njb-49 262 5 , , PUNCT njb-49 262 6 15(12):1819 15(12):1819 NUM njb-49 262 7 - - SYM njb-49 262 8 1823 1823 NUM njb-49 262 9 . . PUNCT njb-49 263 1 35 35 NUM njb-49 263 2 . . PUNCT njb-49 264 1 agarwal agarwal PROPN njb-49 264 2 r r PROPN njb-49 264 3 , , PUNCT njb-49 264 4 binz binz PROPN njb-49 264 5 t t PROPN njb-49 264 6 , , PUNCT njb-49 264 7 swaminathan swaminathan PROPN njb-49 264 8 s s PROPN njb-49 264 9 : : PUNCT njb-49 264 10 structural structural ADJ njb-49 264 11 analysis analysis NOUN njb-49 264 12 of of ADP njb-49 264 13 botulinum botulinum ADJ njb-49 264 14 neurotoxin neurotoxin NOUN njb-49 264 15 serotype serotype VERB njb-49 264 16 f f PROPN njb-49 264 17 light light PROPN njb-49 264 18 chain chain NOUN njb-49 264 19 : : PUNCT njb-49 264 20 implications implication NOUN njb-49 264 21 on on ADP njb-49 264 22 substrate substrate ADJ njb-49 264 23 binding bind VERB njb-49 264 24 and and CCONJ njb-49 264 25 inhibitor inhibitor NOUN njb-49 264 26 design design NOUN njb-49 264 27 . . PUNCT njb-49 265 1 biochemistry biochemistry NOUN njb-49 265 2 2005 2005 NUM njb-49 265 3 , , PUNCT njb-49 265 4 44(35):11758 44(35):11758 NUM njb-49 265 5 - - NUM njb-49 265 6 11765 11765 NUM njb-49 265 7 . . PUNCT njb-49 266 1 36 36 NUM njb-49 266 2 . . PUNCT njb-49 266 3 zdanovsky zdanovsky PROPN njb-49 266 4 ag ag PROPN njb-49 266 5 , , PUNCT njb-49 266 6 zdanovskaia zdanovskaia PROPN njb-49 266 7 mv mv PROPN njb-49 266 8 : : PUNCT njb-49 266 9 simple simple ADJ njb-49 266 10 and and CCONJ njb-49 266 11 efficient efficient ADJ njb-49 266 12 method method NOUN njb-49 266 13 for for ADP njb-49 266 14 heterologous heterologous ADJ njb-49 266 15 expression expression NOUN njb-49 266 16 of of ADP njb-49 266 17 clostridial clostridial ADJ njb-49 266 18 proteins protein NOUN njb-49 266 19 . . PUNCT njb-49 267 1 appl appl VERB njb-49 267 2 environ environ NOUN njb-49 267 3 microbiol microbiol ADJ njb-49 267 4 2000 2000 NUM njb-49 267 5 , , PUNCT njb-49 267 6 66(8):3166 66(8):3166 NUM njb-49 267 7 - - SYM njb-49 267 8 3173 3173 NUM njb-49 267 9 . . PUNCT njb-49 268 1 nepal nepal ADJ njb-49 268 2     SPACE njb-49 268 3 journal journal NOUN njb-49 268 4     SPACE njb-49 268 5 of of ADP njb-49 268 6     SPACE njb-49 268 7 biotechnology biotechnology NOUN njb-49 268 8 . . PUNCT njb-49 268 9         SPACE njb-49 269 1 jan jan PROPN njb-49 269 2 . . PROPN njb-49 269 3     SPACE njb-49 269 4 2012 2012 NUM njb-49 269 5 , , PUNCT njb-49 269 6     SPACE njb-49 269 7 vol vol NOUN njb-49 269 8 . . PUNCT njb-49 269 9     SPACE njb-49 269 10 2 2 NUM njb-49 269 11 , , PUNCT njb-49 269 12     SPACE njb-49 269 13 no no INTJ njb-49 269 14 . . PUNCT njb-49 269 15     SPACE njb-49 270 1 1 1 NUM njb-49 270 2 : : PUNCT njb-49 270 3     SPACE njb-49 270 4 1 1 X njb-49 270 5     SPACE njb-49 270 6 – – PUNCT njb-49 270 7     SPACE njb-49 270 8 15 15 NUM njb-49 270 9                                                                                                                                                                                                                                         SPACE njb-49 270 10 biotechnology biotechnology NOUN njb-49 270 11     SPACE njb-49 270 12 society society NOUN njb-49 270 13     SPACE njb-49 270 14 of of ADP njb-49 270 15     SPACE njb-49 270 16 nepal nepal NOUN njb-49 270 17     SPACE njb-49 270 18 ( ( PUNCT njb-49 270 19 bsn bsn PROPN njb-49 270 20 ) ) PUNCT njb-49 270 21 , , PUNCT njb-49 270 22     SPACE njb-49 270 23 all all DET njb-49 270 24     SPACE njb-49 270 25 rights right NOUN njb-49 270 26     SPACE njb-49 270 27 reserved reserve VERB njb-49 270 28     SPACE njb-49 270 29 15 15 NUM njb-49 270 30 37 37 NUM njb-49 270 31 . . PUNCT njb-49 271 1 woodward woodward PROPN njb-49 271 2 la la PROPN njb-49 271 3 , , PUNCT njb-49 271 4 arimitsu arimitsu PROPN njb-49 271 5 h h PROPN njb-49 271 6 , , PUNCT njb-49 271 7 hirst hirst PROPN njb-49 271 8 r r PROPN njb-49 271 9 , , PUNCT njb-49 271 10 oguma oguma PROPN njb-49 271 11 k k PROPN njb-49 271 12 : : PUNCT njb-49 271 13 expression expression NOUN njb-49 271 14 of of ADP njb-49 271 15 hc hc PROPN njb-49 271 16 subunits subunit NOUN njb-49 271 17 from from ADP njb-49 271 18 c. c. PROPN njb-49 271 19 botulinum botulinum PROPN njb-49 271 20 types type NOUN njb-49 271 21 c c NOUN njb-49 271 22 and and CCONJ njb-49 271 23 d d NOUN njb-49 271 24 and and CCONJ njb-49 271 25 their their PRON njb-49 271 26 evaluation evaluation NOUN njb-49 271 27 as as ADP njb-49 271 28 candidate candidate NOUN njb-49 271 29 vaccine vaccine NOUN njb-49 271 30 antigens antigen NOUN njb-49 271 31 in in ADP njb-49 271 32 mice mouse NOUN njb-49 271 33 . . PUNCT njb-49 272 1 infect infect ADJ njb-49 272 2 immun immun PROPN njb-49 272 3 2003 2003 PROPN njb-49 272 4 , , PUNCT njb-49 272 5 71(5):29412944 71(5):29412944 NUM njb-49 272 6 . . PROPN njb-49 272 7 38 38 NUM njb-49 272 8 . . PUNCT njb-49 273 1 dulal dulal PROPN njb-49 273 2 s s PROPN njb-49 273 3 , , PUNCT njb-49 273 4 ban ban PROPN njb-49 273 5 b b PROPN njb-49 273 6 , , PUNCT njb-49 273 7 kim kim PROPN njb-49 273 8 ks ks PROPN njb-49 273 9 , , PUNCT njb-49 273 10 yang yang PROPN njb-49 273 11 gh gh PROPN njb-49 273 12 , , PUNCT njb-49 273 13 jung jung PROPN njb-49 273 14 hh hh INTJ njb-49 273 15 . . PUNCT njb-49 274 1 : : PUNCT njb-49 274 2 neutralizing neutralize VERB njb-49 274 3 antibody antibody NOUN njb-49 274 4 specific specific ADJ njb-49 274 5 to to ADP njb-49 274 6 c. c. PROPN njb-49 274 7 botulinum botulinum PROPN njb-49 274 8 neurotoxin neurotoxin NOUN njb-49 274 9 serotype serotype VERB njb-49 274 10 f f PROPN njb-49 274 11 ( ( PUNCT njb-49 274 12 bont bont PROPN njb-49 274 13 / / SYM njb-49 274 14 f f NUM njb-49 274 15 ) ) PUNCT njb-49 274 16 . . PUNCT njb-49 275 1 abstracts abstract NOUN njb-49 275 2 of of ADP njb-49 275 3 the the DET njb-49 275 4 110th 110th ADJ njb-49 275 5 general general ADJ njb-49 275 6 meeting meeting NOUN njb-49 275 7 of of ADP njb-49 275 8 the the DET njb-49 275 9 american american ADJ njb-49 275 10 society society PROPN njb-49 275 11 for for ADP njb-49 275 12 microbiology microbiology NOUN njb-49 275 13 ( ( PUNCT njb-49 275 14 american american PROPN njb-49 275 15 society society PROPN njb-49 275 16 for for ADP njb-49 275 17 microbiology microbiology NOUN njb-49 275 18 , , PUNCT njb-49 275 19 san san PROPN njb-49 275 20 diego diego PROPN njb-49 275 21 , , PUNCT njb-49 275 22 ca can AUX njb-49 275 23 , , PUNCT njb-49 275 24 23rd 23rd ADJ njb-49 275 25 27th 27th NOUN njb-49 275 26 may may AUX njb-49 275 27 ) ) PUNCT njb-49 275 28 , , PUNCT njb-49 275 29 p p PROPN njb-49 275 30 110 110 NUM njb-49 275 31 2010 2010 NUM njb-49 275 32 . . PUNCT njb-49 276 1 39 39 NUM njb-49 276 2 . . PUNCT njb-49 276 3 yang yang PROPN njb-49 276 4 gh gh PROPN njb-49 276 5 , , PUNCT njb-49 276 6 kim kim PROPN njb-49 276 7 ks ks PROPN njb-49 276 8 , , PUNCT njb-49 276 9 kim kim PROPN njb-49 276 10 hw hw PROPN njb-49 276 11 , , PUNCT njb-49 276 12 jeong jeong PROPN njb-49 276 13 st st PROPN njb-49 276 14 , , PUNCT njb-49 276 15 huh huh INTJ njb-49 276 16 gh gh PROPN njb-49 276 17 , , PUNCT njb-49 276 18 kim kim PROPN njb-49 276 19 jc jc PROPN njb-49 276 20 , , PUNCT njb-49 276 21 jung jung PROPN njb-49 276 22 hh hh VERB njb-49 276 23 : : PUNCT njb-49 276 24 isolation isolation NOUN njb-49 276 25 and and CCONJ njb-49 276 26 characterization characterization NOUN njb-49 276 27 of of ADP njb-49 276 28 a a DET njb-49 276 29 neutralizing neutralize VERB njb-49 276 30 antibody antibody NOUN njb-49 276 31 specific specific ADJ njb-49 276 32 to to PART njb-49 276 33 internalization internalization VERB njb-49 276 34 domain domain NOUN njb-49 276 35 of of ADP njb-49 276 36 c. c. PROPN njb-49 276 37 botulinum botulinum PROPN njb-49 276 38 neurotoxin neurotoxin PROPN njb-49 276 39 type type PROPN njb-49 276 40 b. b. PROPN njb-49 276 41 toxicon toxicon PROPN njb-49 276 42 2004 2004 NUM njb-49 276 43 , , PUNCT njb-49 276 44 44(1):19 44(1):19 NUM njb-49 276 45 - - SYM njb-49 276 46 25 25 NUM njb-49 276 47 . . NUM njb-49 276 48 40 40 NUM njb-49 276 49 . . PUNCT njb-49 277 1 baldwin baldwin PROPN njb-49 277 2 mr mr PROPN njb-49 277 3 , , PUNCT njb-49 277 4 tepp tepp PROPN njb-49 277 5 wh wh PROPN njb-49 277 6 , , PUNCT njb-49 277 7 przedpelski przedpelski VERB njb-49 277 8 a a PRON njb-49 277 9 , , PUNCT njb-49 277 10 pier pier NOUN njb-49 277 11 cl cl PROPN njb-49 277 12 , , PUNCT njb-49 277 13 bradshaw bradshaw PROPN njb-49 277 14 m m PROPN njb-49 277 15 , , PUNCT njb-49 277 16 johnson johnson PROPN njb-49 277 17 ea ea PROPN njb-49 277 18 , , PUNCT njb-49 277 19 barbieri barbieri PROPN njb-49 277 20 jt jt PROPN njb-49 277 21 : : PUNCT njb-49 277 22 subunit subunit PROPN njb-49 277 23 vaccine vaccine NOUN njb-49 277 24 against against ADP njb-49 277 25 the the DET njb-49 277 26 seven seven NUM njb-49 277 27 serotypes serotype NOUN njb-49 277 28 of of ADP njb-49 277 29 botulism botulism NOUN njb-49 277 30 . . PUNCT njb-49 278 1 infect infect PROPN njb-49 278 2 immun immun PROPN njb-49 278 3 2008 2008 PROPN njb-49 278 4 , , PUNCT njb-49 278 5 76(3):1314 76(3):1314 NUM njb-49 278 6 - - SYM njb-49 278 7 1318 1318 NUM njb-49 278 8 . . PUNCT njb-49 279 1 41 41 NUM njb-49 279 2 . . PUNCT njb-49 279 3 webb webb PROPN njb-49 279 4 rp rp PROPN njb-49 279 5 , , PUNCT njb-49 279 6 smith smith PROPN njb-49 279 7 tj tj PROPN njb-49 279 8 , , PUNCT njb-49 279 9 wright wright PROPN njb-49 279 10 pm pm PROPN njb-49 279 11 , , PUNCT njb-49 279 12 montgomery montgomery PROPN njb-49 279 13 va va PROPN njb-49 279 14 , , PUNCT njb-49 279 15 meagher meagher PROPN njb-49 279 16 mm mm PROPN njb-49 279 17 , , PUNCT njb-49 279 18 smith smith PROPN njb-49 279 19 la la PROPN njb-49 279 20 : : PUNCT njb-49 279 21 protection protection NOUN njb-49 279 22 with with ADP njb-49 279 23 recombinant recombinant ADJ njb-49 279 24 c. c. PROPN njb-49 279 25 botulinum botulinum PROPN njb-49 279 26 c1 c1 PROPN njb-49 279 27 and and CCONJ njb-49 279 28 d d X njb-49 279 29 binding bind VERB njb-49 279 30 domain domain NOUN njb-49 279 31 subunit subunit NOUN njb-49 279 32 ( ( PUNCT njb-49 279 33 hc hc PROPN njb-49 279 34 ) ) PUNCT njb-49 279 35 vaccines vaccine NOUN njb-49 279 36 against against ADP njb-49 279 37 c c PROPN njb-49 279 38 and and CCONJ njb-49 279 39 d d PROPN njb-49 279 40 neurotoxins neurotoxin NOUN njb-49 279 41 . . PUNCT njb-49 280 1 vaccine vaccine PROPN njb-49 280 2 2007 2007 NUM njb-49 280 3 , , PUNCT njb-49 280 4 25(21):4273 25(21):4273 NUM njb-49 280 5 - - SYM njb-49 280 6 4282 4282 NUM njb-49 280 7 . . PUNCT njb-49 281 1 42 42 NUM njb-49 281 2 . . PUNCT njb-49 281 3 webb webb PROPN njb-49 281 4 rp rp PROPN njb-49 281 5 , , PUNCT njb-49 281 6 smith smith PROPN njb-49 281 7 tj tj PROPN njb-49 281 8 , , PUNCT njb-49 281 9 wright wright NOUN njb-49 281 10 p p PROPN njb-49 281 11 , , PUNCT njb-49 281 12 brown brown PROPN njb-49 281 13 j j PROPN njb-49 281 14 , , PUNCT njb-49 281 15 smith smith PROPN njb-49 281 16 la la PROPN njb-49 281 17 : : PUNCT njb-49 281 18 production production NOUN njb-49 281 19 of of ADP njb-49 281 20 catalytically catalytically PROPN njb-49 281 21 inactive inactive ADJ njb-49 281 22 bont bont PROPN njb-49 281 23 / / SYM njb-49 281 24 a1 a1 PROPN njb-49 281 25 holoprotein holoprotein PROPN njb-49 281 26 and and CCONJ njb-49 281 27 comparison comparison NOUN njb-49 281 28 with with ADP njb-49 281 29 bont bont PROPN njb-49 281 30 / / SYM njb-49 281 31 a1 a1 PROPN njb-49 281 32 subunit subunit NOUN njb-49 281 33 vaccines vaccine NOUN njb-49 281 34 against against ADP njb-49 281 35 toxin toxin NOUN njb-49 281 36 subtypes subtype NOUN njb-49 281 37 a1 a1 PROPN njb-49 281 38 , , PUNCT njb-49 281 39 a2 a2 PROPN njb-49 281 40 , , PUNCT njb-49 281 41 and and CCONJ njb-49 281 42 a3 a3 PROPN njb-49 281 43 . . PUNCT njb-49 282 1 vaccine vaccine PROPN njb-49 282 2 2009 2009 NUM njb-49 282 3 , , PUNCT njb-49 282 4 27(33):4490 27(33):4490 NUM njb-49 282 5 - - SYM njb-49 282 6 4497 4497 NUM njb-49 282 7 . . PUNCT