





































Clinical Medicine Insights
Received 10 Feb 2022 | Revised 13 Feb 2022 | Accepted 18 Feb 2022 | Published Online 20 Feb 2022

DOI: https://doi.org/10.52845/CMI/2022-3-1-4
CMI 02 (02), 262−267 (2022) ISSN (O) 2694-4626 IF:1.6

RESEARCH ARTICLE

Vaccines against potential pathogens[Vaccine against Plague. ]

DR.Michael Vladislavovich Tyurin M.D., Ph.D
1CEO, Executive Chairman,
President of Microbial Biocatalyst
International, Inc. and
Inorgcarbdiesel, Inc. Dr. Tyurin
has gained two Doctoral Degrees
in the former Soviet Union: M.D.
in Internal Medicine (1986,
Saratov State Medical University)
and Ph.D. in Molecular Biology,
Microbiology and Molecular
Pharmacology (1990, The USSR
Research Institute for Antibiotics).
Dr. Tyurin additionally has B. Sci.
in Biological Engineering (1984)
and M. Sci. in Biology (1985) both
from Saratov State Medical
University.

Abstract
We proposed human intestine as the gate for the delivery of the 
therapeutic recombinant proteins expressed inside of the human body. 
The normal intestinal microflora was used to express the selected genes 
of the pathogenic organism Yersinia pestis the causative agent for plug 
in humans and animals. We have confirmed the production of the 
selected proteins by the PCR to their DNAs expressed in intestinal 
bifidobacteria chosen. Now it is the role of the immunologists to find 
the antibodies for the recombinant proteins expressed inside of the 
volunteer’s organism.
Keywords:   Expression of the recombinant proteins inside human’s 
body, Bifidobacterium breve, use of genome tailoring technology to 
express the recombinant proteins inside of the volunteer’s body.

Copyright : © 2022 The Authors. Published by Publisher. This is an
open access article under the CC BY-NC-ND license
(https://creativecommons.org/licenses/by-nc-nd/4.0/).

1 INTRODUCTION

The Author started his medical education at
Saratov State Medical University after he
spoke with his cousin Galina about becoming

the Ph.D-Scientist working with the Yesrinia pestis
the causative agent of plague and Vibrio cholera
the causative agent of cholera at the closed for
the employment of the general public Institution
Microbe in Author’s native city Saratov (now dis-
solved). Said scientists had salaries substantially ex-
ceeding that of the regular former Soviet society
members, just like the Author. For instance, the
Soviet Academician was getting his salary of 1,000
Russian rubles, while at the mentioned organization

Microbe Senior Ph.D. Researcher was getting 2,600
rubles, etc. The Author has made multiple friends
among said Ph.D-.level Scientists from that closed
for general public employment institution Microbe.
Some contacts became very useful for Dr. Tyurin
for making his scientific presentations at Saratov
State Medical University during his course of study-
ing. The most remarkable was the gift of the Mi-
crobe Institute medical advisor when she gave to
Dr. Tyurin 5 kg of Japanese Agar-Agar. The Author
could use that agar during his already scheduled
Ph.D. studies at the USSR Research Institute for
Antibiotics inMoscow, the former USSR, and for his
future work as the Ph.D-student in Moscow, when
Dr. Tyurin has gotten as a present those 5 kg of

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MANUSCRIPT CENTRAL
M.D., PH.D

the Japanese Agar-Agar he later used in Moscow
for his Ph.D-associated research and development
(R & D). While studying at Saratov State Medical
University and visiting the Microbiology and Im-
munology Department of said University from early
in 1981 till his graduation date in 1986 the Author
has learned that what he wanted to become after the
graduation of Saratov State Medical University was
not possible, complicated by his origin of the regular
private person. He must note that his cousin Galina
was the daughter of the SPCU (Soviet Communist
Party) Second Secretary of the Saratov Region SPCU
Committee Vladimir Rodionov. That gave her the
protected path to become the Ph.D-Scientist in said
closed for the general public employment former So-
viet Union institution Microbe dealing with plague-
cholera causative agents during the futile scientists
attempts to create the new biological weapons of
mass destruction with the tremendous destructive
power and difficulty to cure. That mentioned closed
system required mandatory thorough checking of the
background by the KGB and the origination from
families of the high ranked SPCU-related people
in the former Soviet Union, which was quite op-
posite to what the Author had in his background.
While learning about that in the course of study at
Saratov State Medical University, the Author con-
tinued to want to become the professional Ph.D.
level scientist working with microorganisms, and he
was not any longer inspired by the levels of the
respective salaries such noted secretly working on
the weapons of the mass destruction Ph.D.-Scientists
had in the former Soviet Union. On the second year
of the Author’s education (1982) at Saratov State
Medical University the Author has met Professor
Boris Shenderov, a person who just returned from
Zambia where he worked as the Professor at the
Lusaca University. Dr. Shenderov worked for the
KGB and that was the reason he was in Zambia
for his “work”. Dr. Shenderov visited Saratov State
Medical University he graduated from as well. He
has visited Saratov State Medical University in 1984
before moving to Moscow, where the former Soviet
KGB gave Dr. Shenderov the rank of the colonel
and the new work of the Professor at the Laboratory
of the Industrial Hygiene at the USSR Research
Institute for Antibiotics, Moscow, the former USSR.

So, Dr. Shenderov used data the Author has provided
to him on the Author’s studies of Non-Fermenting
Glucose Gram Negative organisms predominantly
Pseudomonas isolated from the hospital patients in
the Saratov Region. Said data gave the opportunity
to publish them in the Journal for Dr. Shenderov’s
future Moscow Institution “Antibiotics” in 1984 (the
1st publication Dr, Tyurin had in the Former Soviet
Union [1] while being a simple medical student).
Dr. Shenderov invited Dr. Tyurin to continue the
education in Moscow to become his Ph.D-Student
upon the graduation of the Saratov State Medical
University, which the Author did in 1986. Upon
the graduation of his PhD.-Studentship the Author
joined Dr. Shenderov at his another new work place,
as Dr. Shenderov got the promotion from the KGB
for his work in Moscow since 1984, at Gabrichevsky
Research Institute for Epidemiology and Microbiol-
ogy in 1990 where Dr Shenderov became the Di-
rector. In 1992 the Author has left Dr. Shenderov
and his KGB-related work community and joined
theAuthor’s newwork acquired at VNIIGENETIKA
(the Adjinomoto-GNIIGENETIKA Research Insti-
tute), the Author’s last place of work at the Russian
Federation before moving permanently to the USA.
The Author has already described the known before
place of entry to the human body bloodstream -
human intestine [2,3]. The Author had specialization
during his Ph.D.-Studentship years on the Molecular
Biology, Microbiology and Molecular Pharmacol-
ogy of the normal intestinal microflora of humans
and animals. The Author has become the king of
lactobacilli and substantially intensified his work
with the human intestinal bifidobacteria at GNI-
IGENETIKA in 1992-1998, the predominant organ-
isms in the intestine of many humans [4]. In this orig-
inal research paper the Author describes his personal
experience with the volunteer he has successfully
vaccinated by creating the recombinant strain of
bifidobacteria isolated from the intestinal content of
said volunteer as described in [2,3] and making said

Supplementary information The online version
of this article (10.52845/CMI/2022-3-1-4) contains
supplementary material, which is available to autho-
rized users.

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VACCINES AGAINST POTENTIAL PATHOGENS[VACCINE AGAINST PLAGUE. ]

volunteer to ingest the recombinant strain of his own
intestinal bifidobacteria.
As the target for the expression in the recombi-
nant Bifidobacterium breve 839 strain the Author
has chosen the DNA with the known nucleic acid
content originally isolated from the Yersinia pestis
strain. Said strain was the causative agent for the
human and animals plug [6]. The Author’s choice
for said proteins was dictated by the ethiological
role of the Yersinia pestis selected as the possible
ethiologic agent of the emergent diseases the outer
Space travel crews might possibly face discovering
other new planets similar to Earth by the temperature
and the atmosphere content in the coming future, the
immediate projects of the Environmental direction. .

2 MATERIALS AND METHODS

The isolation and investigation of intestinal bifi-
dobacteria of the volunteer was performed as de-
scribed [2,3]. Using the described selective medium
for the isolation of the intestinal bifidobacteria
[2] the Author has isolated Bifidobacterium breve
839 strain from said volunteer’s freshly collected
intestinal content (fresh feces). Said strain was
subjected to the reduction of its genome by removal
of not essential for the vital functions of said
strain natural genome genes at their positions
4346...4816 bp,10023...10574 bp, 16239...17477 bp,
19324...20316 bp, 20927...21592 bp,22486...23799
bp,237007...238836 bp, 24676...26007 bp,
31295...33502 bp,34410...36290 bp, 37707...39254
bp, 538583...541012 bp, 643441...645516 bp,
817368...820625 bp, 1476554...1481404 bp and
2258202...2261981 bp using the procedures de-
scribed in [7-16].
Total genomic DNA from B. breve 839 was isolated
by the procedure [4,18]. The primers for the PCR
to check the presence of the recombinant pesticin
and the hypothetical protein YPMT1.21c DNA se-
quences were designed using the publically available
tool [17].
The process of genetic modification of said B. breve
839 strain took less then 200 hours to ensure the
strain regained the capability to adhere back to the

intestinal wall of the volunteer as we have discussed
before [2,3].
Bifidobactrium breve 839 pesticin Submission ID
2526713.
Bifidobacterium breve 839 hypotetical protein
YPMT1.21c submission ID 2532183.

3 RESULTS

Development of the recombinant strain of B. breve
839 PLUG was performed as already described in
[2,3]. Said strain of intestinal human bifidobacte-
ria of the volunteer has expressed both recombi-
nant genes of pesticin and the hypothetical protein
YPMT1.21c as that was shown by the PCR the
Author has performed with the total genomic DNA
isolated from B. breve 839 PIUG by the method
described in [18] with the proper modifications of the
procedure to isolate the total genomic DNA.
The PCR for the presence of the both recombi-
nant genes of pesticin and the hypothetical protein
YPMT1.21c ended up with the revealing of the
proper DNA fragments on the agarose gel for the
PCR products [17]. Upon said testing the 25ml of the
72-h broth culture of B. breve 839 PLUG grown in
the liquid m3edium for the isolation of bifidobacteria
with no antibiotics added were. ingested by the vol-
unteer. In one hour he came back to his work and has
ever worked as before the ingestion. Now, it is the
task for the immunologists to investigate the blood
stream of said volunteer to recover the antibodies
to the Yersinia pestis pesticin and the hypotetical
protein YPMT1.21c.

4 DISCUSSION

We have discussed the prospects of our planet in
the future at our corporate web site, and noted the
coming in 10-20 years from now the shortage of the
fresh water, leaving the Earth to the outer Space as
described [2,3]. Indeed, accumulated in the air CO2 is
one of the heaviest gasses in the air blend, reaching
its density 1.97 g/cubic meter [3]. The CO2 in the
air gas mixture under the no wind environmental

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M.D., PH.D

conditions spreads on the ground surface and selec-
tively absorbs all the infrared energy of the Sun light,
thus heating the ground significantly. That causes
the extra evaporation of the fresh water from soil
to the air. As you know, Global Warming presents
itself in various forms, specifically with increased
frequency of rainy weather, long rainy days, tor-
nadoes, etc. But the Earth gravity has been stable
for the last few million years from now. Therefore,
under the constant gravity force applied, more fresh
water vapors are in the air. The space, surrounding
Earth, as any Space anywhere, has vacuum. That
vacuum sucks fresh water vapors from Earth air, and
such fresh water vapors travel in the Space in the
unknown direction away from the Earth. In 2010
NASA has bombarded the Moon and found plenty
of ice on its dark and very cold surface. The Earth
satellite Moon is located 220,000 miles away from
Earth. One Moon’s side is always dark and cold as
it never gets Sun light irradiation. It is very cold, as
cold as the Space vacuum, -273 o C. So NASA were
guessing where said ice came from? Moon worked
as the cold trap for the fresh water vapors coming
from Earth in the Space vacuum [http://syngasbio
fuelsenergy.com]. What will happen next and the
most important, when?Wemight give the time frame
for 10-20 or 10-50 years from now, based on the
2010 HASA discovery of ice on the Moon and the
NASA conclusion of their findings: Earth as planet
has passed the “point of no return” to the normal
life. It is impossible to anticipate, that the fresh water
loss to the outer Space may be stopped at any time
even if the Earth population is suddenly decreased
in its amount. The extra air CO2 comes from the
intensified petroleum use, and the use of its products
for combustion, producing CO2. People breath and
produce CO2 as well. It is anticipated the 14 billion
people on Earth by 2050. That increases more the air
CO2 content, leading to the increased freshwater loss
as discussed.

Fig 1 shows our understanding of the fresh water loss
process.

Fig, 1. Fresh water loss to the Space vacuum
We have no any idea, what will happen soon, if
no new planets, similar to Earth, will be discovered
and the overcrowded Earth population will not start
to move there. We do anticipate, that the reduction
of the air CO2 content is absolutely necessary and
possible by the replacing of the existing economy
based on the power generation suing the products
of petroleum distillation on the petroleum refineries
by the economy based on the energy generation
using the discovered by the Author carbon negative
technologies of the fuels and chemicals production
[2-16].
In this article, the Author has shown that the ex-
pression of the recombinant proteins originated from
Yersinia pestis and expressed in the human intestinal
bifidobacteria happens efficiently right in the body
of the volunteer by the engineered strain of the
intestinal bifidobacterium isolated and then returned
back to the intestine of said volunteer. Based on the
described expression of the recombinant proteins,
the Author has concluded, that that will be possible
to perform the immunization of the proposed com-
ing soon manned crews of the outer Space flights
intended to discover new planets in the Universe
suitable for the relocation of the overcrowded Earth.
Said immunization does require certain genetic ma-
nipulations which are possible on the board of said
outer Space travel vehicle(s) by the suing the de-
scribed technologies of genome replacement in the
intestinal microflora of the potential crew members
of said vehicle. This circumstance closes the need for

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VACCINES AGAINST POTENTIAL PATHOGENS[VACCINE AGAINST PLAGUE. ]

the special medical personnel on board of said outer
Space travel vehicle(s) to perform the immunization
of the crew from the emergent infections reasonably
anticipated for the existence in the new outer Space
locations. This approach may have crucial impor-
tance for the manned crews life during said long term
outer space travel missions proposed.

5 DECLARATIONS

Ethical Approval and Consent to Participate. This
article does not contain any section, requiring Ethical
Approval. The only Author is complied with the
Consent to Participate. Consent to Participate. The
Author complied with the consent to publish this
article.
Consent to Publish. This original article has not
been published anywhere or is under the consid-
eration to publish anywhere else beside this Jour-
nal. The Author complied with the Consent to Pub-
lish this original article. Authors Contribution. Dr.
Michael V. Tyurin has planned all the experimental
work, conducted all the experiments, analyzed the
experimental data, wrote this original manuscript,
edited ot as appropriate and submitted for this pub-
lication the edited original manuscript. FUNDING
was done by the private investors, who declined
to provide their names and their business affilia-
tions. The investors noted, the author should de-
cline any source of funding. Competing Interests.
The author declares his personal conflict of interests
with the law firm in Houston, TX Hirsch and Wes-
theimer, which has destroyed his corporate website
http:syngasbiofuelsenergy.com, and with the major
petroleum and gasoline/diesel fuel companies in
Houston (TX), with the Houston Police (City of
Houston) and with the Houston FBI ignoring the
Author’s concern about his attempted murder com-
mitted by SHELL after the Author has presented
to SHELL his proprietary technology for gasoline
manufacture from the air CO2, not from petroleum
(Tyurin MV, et al. 2019) The attempted murder is
of no interest for the corrupt by the major petroleum
corporations in Houston, TX Houston FBI. The Au-
thor has no intent to file the lawsuit against the
Houston FBI at any point, but he is inclined to make

this case the public domain. Houston FBI does not
follow the established in the USA Federal Laws.
Availability of data and materials, All the data and
materials are available if necessary from the Au-
thor of this manuscript. The Authors’ information.
The Author is the owner of his mentioned TEXAS
businesses Microbial Biocatalyst International, Inc.
and Inorgcarbdiesel, Inc. The work has been done at
the corporate site with the USPS address P. O. Box
300230, Houston, TX 77230.

6 REFERENCES

1. Shenderov BA, Serkova GP, Tyurin MV (1984)
Susceptibility of clinical non-fermenting Gram-
Negative bacteria to antibacterial drugs // Antibiotiki
29 (3):191–195.
2. Tyurin MV (2021) Expression in situ of the
Recombinant Human Erythropoetin and Recom-
binant Insulin. J Diabetes Metab. 12:900. doi:
10.35248/2252-5211.21.12.900
3. Tyurin MV (2021) Successful Treatment of Dia-
betes II in adult patient and New Prospects of Re-
combinant Vaccine and Recombinant Proteins Engi-
neering in situ. J Diabetes Metab. 12:871-875.
4. Tyurin M.V. Ph.D. - Thesis: ”Antibiotic Re-
sistance and Antagonistic Activity of Lactobacilli”
1990, The USSR Research Institute for Antibiotics,
Moscow, the USSR.
5. http://syngasbiofuelsenergy.com.
6. ParkhillJ,Wren B.W.,Thomson N.R.,Titball
R.W.,Holden M.T.,Prentice M.B.,Sebaihia
M.,James K.D.,Churcher C.,Mungall K.L.,Baker
S.,Basham D.,Bentley S.D.,Brooks K.,Cerdeño-
Tárraga A.M.,Chillingworth T.,Cronin A.,Davies
R.M.,Davis P.,Dougan G.,Feltwell T.,Hamlin
N.,Holroyd S.,Jagels K.,Karlyshev A.V.,Leather
S.,Moule S.,Oyston P.C.,Quail M.,Rutherford
K.,Simmonds M.,Skelton J.,Stevens K.,Whitehead
S.,Barrel B.G. (2001).Genome sequence of Yersinia
pestis, the causative agent of plague. Nature. 2001
Oct 4; 413 (6855): 523-7. doi: 10.1038/35097083.
7. Tyurin M, Kiriukhin M (2013). 2,3-Butanediol
production by engineered acetogen biocatalyst dur-
ing continuous fermentation of syngas or CO2/H2

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M.D., PH.D

blend. Appl
8. Tyurin M, Kiriukhin M. (2013). Selective
methanol or formate production during continuous
CO2fermentation by the acetogen biocatalysts
engineered via integration of synthetic pathways
using Tn7-tool. World Journal of Microbiol-
ogy and Biotechnology. 29 (9)1611-1623. doi:
10.1007/s11274-013-1324-2.
9. Tyurin M. (2013). Gene replacement and elimi-
nation using λRed- and FLP-based tool to re-direct
carbon flux in acetogen biocatalyst during continu-
ous CO2/H2 blend fermentation. Journal of Industrial
Microbiology&Biotechnology. 40 (7):749-758. doi:
10.1007/s10295-013-1279-1.
10. Berzin V, Kiriukhin M, Tyurin M. (2012)
Selective production of acetone during continu-
ous synthesis gas fermentation by engineered bio-
catalyst Clostridium sp. MAceT113. Letters of
Appl Microbiol. 55(2):149-54. doi: 10.1111/j.1472-
765X.2012.03272.x.
11. Tyurin M, Kiriukhin M. (2013). Expression of
amplified synthetic ethanol pathway integrated using
Tn7-tool and powered at the expense of eliminated
pta, ack, spo0A and spo0J during continuous syngas
or CO2 /H2 blend fermentation. J Appl Microbiol.
114(4):1033-45. doi: 10.1111/jam.12123.
12. Tyurin M, Kiryukhin M, Berzin V. (2012) Elec-
trofusion of untreated cells of the newly isolated ace-
togen Clostridium sp. MT351 with integrated in the
chromosome erm(B) or cat leading to the combined
presence of these antibiotic resistance genes 1in the
chromosome of the electrofusion products. Journal
of Biotech Research. 4:1-12.
13. Berzin V, Kiriukhin M, Tyurin M. (2013)
Cre-lox66/lox71-based elimination of phospho-

transacetylase or acetaldehyde dehydrogenase
shifted carbon flux in acetogen rendering selective
overproduction of ethanol or acetate. Appl Biochem
Biotechnol. 195(3):181-8. http://www.ncbi.nlm.nih.
gov/pubmed/22941272.
14. Berzin V, Kiriukhin M, Tyurin M. (2013) Se-
lective n-butanol production by Clostridium sp.MT-
ButOH1365 during continuous synthesis gas fer-
mentation due to expression of synthetic thiolase,
3-hydroxy butyryl-CoA dehydrogenase, crotonase,
butyryl-CoA dehydrogenase, butyraldehyde dehy-
drogenase and NAD-dependent butanol dehydroge-
nase. Appl Biochem Biotechnol. 169(3), 950-959.
doi: 10.1007/s12010-012-0060-7.
15. Bifidobacterium breve. Normal gastrointestinal
bacterium. 1855. Begey’s Manual of Determinative
Bacteriology, Ninth Edition.
16. Tyurin MV, Padda RS. Nitrogen gas reducing
commercial acetogen biocatalyst suitable for direct
and selective reduction of CO2 inorganic carbon
to organic carbon and atmospheric nitrogen to fuel
isobutanol during continuous fermentation of CO2 +
H2 + N2 gas blend. IRJASET. 2019. 3:1-10.
17. Primer3_results.cgi release 0.4.0.
18. Tyurin MV, Donskih EE, Shenderov BA, Gon-
charova GI (1990) Plasmid DNA isolation method
for bifidobacteria. Antibiot Khimioter 35 (7): 21–22.

How to cite this article: M.D., Ph.D D.R.M.V.T.
Vaccines against potential pathogens[Vaccine
against Plague. ]. Clinical Medicine Insights.
2022;262−267. https://doi.org/10.52845/CMI/2022-
3-1-4

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	Introduction
	Materials and Methods
	Results
	Discussion
	DECLARATIONS 
	References

