Avirulent Probiotic Strain of Escherichia coli, Producing the B-Subunit of Cholera Toxin: Obtainment, Stability of Inheritance, and Expression of the Cloned ctxB Gene
https://doi.org/10.21055/0370-1069-2025-4-156-162
Abstract
The aim of the study was to obtain a cholera toxin B-subunit producer based on the avirulent probiotic strain Escherichia coli M-17 and to study its molecular-genetic and immunogenic properties.
Materials and methods. The genetically marked E. coli KS164 thy polA (pIEM3) strain, containing a recombinant plasmid with the cloned ctxB gene encoding the B-subunit of cholera toxin, and the avirulent E. coli M-17 strain, applied for the production of colicontaining probiotics, were used. The strains were analyzed using various methods: molecular genetics, biochemistry, and immunology.
Results and discussion. Through introducing the recombinant cointegrate plasmid pIEM3 (KmRTcR) with the cloned ctxB gene into E. coli M-17 cells via conjugation, KmRTcR transconjugants with high levels of secreted cholera toxin B-subunit production (10 μg/ml) were obtained. One of these transconjugants (KM147) was used as the producer strain for the protective antigen. It was demonstrated that the constructed E. coli strain KM147(pIEM3) KmRTcR stably inherits the cloned ctxB gene contained in the recombinant plasmid, is capable of producing antitoxic antibodies in immunized animals, and safe for the use in model animals. These properties combined make it promising for future use in the development of prophylactic and diagnostic cholera drugs.
About the Authors
G. V. Chekhovskaya
Russian Research Anti-Plague Institute “Microbe”
Russian Federation
Russian Federation
Galina V. Chekhovskaya,
46, Universitetskaya St., Saratov, 410005
I. M. Krepostnova
Russian Research Anti-Plague Institute “Microbe”
Russian Federation
Russian Federation
46, Universitetskaya St., Saratov, 410005
D. A. Rybal’chenko
Russian Research Anti-Plague Institute “Microbe”
Russian Federation
Russian Federation
46, Universitetskaya St., Saratov, 410005
E. Yu. Shchelkanova
Russian Research Anti-Plague Institute “Microbe”
Russian Federation
Russian Federation
46, Universitetskaya St., Saratov, 410005
N. I. Smirnova
Russian Research Anti-Plague Institute “Microbe”
Russian Federation
Russian Federation
46, Universitetskaya St., Saratov, 410005
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Review
For citations:
Chekhovskaya G.V., Krepostnova I.M., Rybal’chenko D.A., Shchelkanova E.Yu., Smirnova N.I. Avirulent Probiotic Strain of Escherichia coli, Producing the B-Subunit of Cholera Toxin: Obtainment, Stability of Inheritance, and Expression of the Cloned ctxB Gene. Problems of Particularly Dangerous Infections. 2025;(4):156-162. (In Russ.) https://doi.org/10.21055/0370-1069-2025-4-156-162
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