{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,16]],"date-time":"2026-03-16T11:36:24Z","timestamp":1773660984591,"version":"3.50.1"},"reference-count":72,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T00:00:00Z","timestamp":1635379200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Centre for Strategic Planning of FMBA","award":["0373100122120000010"],"award-info":[{"award-number":["0373100122120000010"]}]},{"name":"Fundamental Research Program for the State Academies of Sciences","award":["\u0410\u0410\u0410\u0410-\u041019-119010590015-8"],"award-info":[{"award-number":["\u0410\u0410\u0410\u0410-\u041019-119010590015-8"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Approximately 1\/6 of humanity is at high risk of experiencing cholera epidemics. The development of effective and safe vaccines against Vibrio cholerae, the primary cause of cholera, is part of the public health measures to prevent cholera epidemics. Natural nontoxigenic V. cholerae isolates represent a source of new genetically improved and relatively safe vaccine strains. However, the genomic engineering of wild-type V. cholerae strains is difficult, and these strains are genetically unstable due to their high homologous recombination activity. We comprehensively characterized two V. cholerae isolates using genome sequencing, bioinformatic analysis, and microscopic, physiological, and biochemical tests. Genetic constructs were Gibson assembled and electrotransformed into V. cholerae. Bacterial colonies were assessed using standard microbiological and immunological techniques. As a result, we created a synthetic chromoprotein-expressing reporter operon. This operon was used to improve the V. cholerae genome engineering approach and monitor the stability of the genetic constructs. Finally, we created a stable candidate V. cholerae vaccine strain bearing a recA deletion and expressing the \u03b2-subunit of cholera toxin. Thus, we developed a strategy for the rapid creation of genetically stable and relatively safe candidate vaccine strains. This strategy can be applied not only to V. cholerae but also to other important human bacterial pathogens.<\/jats:p>","DOI":"10.3390\/ijms222111657","type":"journal-article","created":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T23:51:36Z","timestamp":1635465096000},"page":"11657","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Strategy for the Rapid Development of a Safe Vibrio cholerae Candidate Vaccine Strain"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5203-0787","authenticated-orcid":false,"given":"Dmitry S.","family":"Karpov","sequence":"first","affiliation":[{"name":"Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov str., 32, 119991 Moscow, Russia"},{"name":"Bach Institute of Biochemistry, Federal Research Centre \u201cFundamentals of Biotechnology\u201d of the Russian Academy of Sciences, 119071 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna V.","family":"Goncharenko","sequence":"additional","affiliation":[{"name":"Bach Institute of Biochemistry, Federal Research Centre \u201cFundamentals of Biotechnology\u201d of the Russian Academy of Sciences, 119071 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Evgenii V.","family":"Usachev","sequence":"additional","affiliation":[{"name":"Bach Institute of Biochemistry, Federal Research Centre \u201cFundamentals of Biotechnology\u201d of the Russian Academy of Sciences, 119071 Moscow, Russia"},{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1965-0700","authenticated-orcid":false,"given":"Daria V.","family":"Vasina","sequence":"additional","affiliation":[{"name":"Bach Institute of Biochemistry, Federal Research Centre \u201cFundamentals of Biotechnology\u201d of the Russian Academy of Sciences, 119071 Moscow, Russia"},{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elizaveta V.","family":"Divisenko","sequence":"additional","affiliation":[{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7901-5226","authenticated-orcid":false,"given":"Yaroslava M.","family":"Chalenko","sequence":"additional","affiliation":[{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1107-9351","authenticated-orcid":false,"given":"Andrei A.","family":"Pochtovyi","sequence":"additional","affiliation":[{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"},{"name":"Department of Virology, Biological Faculty, Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6634-9879","authenticated-orcid":false,"given":"Roman S.","family":"Ovchinnikov","sequence":"additional","affiliation":[{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valentin V.","family":"Makarov","sequence":"additional","affiliation":[{"name":"Centre for Strategic Planning of FMBA of Russia, 119121 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sergei M.","family":"Yudin","sequence":"additional","affiliation":[{"name":"Centre for Strategic Planning of FMBA of Russia, 119121 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Artem P.","family":"Tkachuk","sequence":"additional","affiliation":[{"name":"Bach Institute of Biochemistry, Federal Research Centre \u201cFundamentals of Biotechnology\u201d of the Russian Academy of Sciences, 119071 Moscow, Russia"},{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9397-3762","authenticated-orcid":false,"given":"Vladimir A.","family":"Gushchin","sequence":"additional","affiliation":[{"name":"Bach Institute of Biochemistry, Federal Research Centre \u201cFundamentals of Biotechnology\u201d of the Russian Academy of Sciences, 119071 Moscow, Russia"},{"name":"N.F. Gamaleya Federal Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Gamaleya str., 18, 123098 Moscow, Russia"},{"name":"Department of Virology, Biological Faculty, Lomonosov Moscow State University, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1539","DOI":"10.1016\/S0140-6736(17)30559-7","article-title":"Cholera","volume":"390","author":"Clemens","year":"2017","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"A118","DOI":"10.1016\/j.vaccine.2019.12.011","article-title":"Current and future cholera vaccines","volume":"38","author":"Shaikh","year":"2020","journal-title":"Vaccine"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1126\/science.aad5901","article-title":"Genomic history of the seventh pandemic of cholera in Africa","volume":"358","author":"Weill","year":"2017","journal-title":"Science"},{"key":"ref_4","unstructured":"World Health Organization (2010). 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