A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain
Approximately 1/6 of humanity is at high risk of experiencing cholera epidemics. The development of effective and safe vaccines against <i>Vibrio cholerae,</i> the primary cause of cholera, is part of the public health measures to prevent cholera epidemics. Natural nontoxigenic <i>...
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| Autores principales: | , , , , , , , , , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/796c770d9b3442c18ad01745fc78b4dc |
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| Sumario: | Approximately 1/6 of humanity is at high risk of experiencing cholera epidemics. The development of effective and safe vaccines against <i>Vibrio cholerae,</i> the primary cause of cholera, is part of the public health measures to prevent cholera epidemics. Natural nontoxigenic <i>V. cholerae</i> isolates represent a source of new genetically improved and relatively safe vaccine strains. However, the genomic engineering of wild-type <i>V. cholerae</i> strains is difficult, and these strains are genetically unstable due to their high homologous recombination activity. We comprehensively characterized two <i>V. cholerae</i> isolates using genome sequencing, bioinformatic analysis, and microscopic, physiological, and biochemical tests. Genetic constructs were Gibson assembled and electrotransformed into <i>V. cholerae</i>. 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 <i>V. cholerae</i> genome engineering approach and monitor the stability of the genetic constructs. Finally, we created a stable candidate <i>V. cholerae</i> vaccine strain bearing a <i>recA</i> deletion and expressing the β-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 <i>V. cholerae</i> but also to other important human bacterial pathogens. |
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