Intracellular Expression of CTB in Vibrio cholerae Strains in Laboratory Culture Conditions

Choi Hunseok; Son Seonghyeon; Lee Donghyun; Bae Jonghyun; Seo Eunyoung; Kim Dong Wook; Kim Eun Jin

doi:10.4014/jmb.2302.02014

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자료유형: 학술저널

저자정보: Choi Hunseok (Department of Pharmacy College of Pharmacy Hanyang University Ansan 15588 Republic of KoreaInstitute of Pharmacological Research Hanyang University Ansan 15588 Republic of Korea) Son Seonghyeon (Department of Pharmacy College of Pharmacy Hanyang University Ansan 15588 Republic of KoreaInstitute of Pharmacological Research Hanyang University Ansan 15588 Republic of Korea) Lee Donghyun (Department of Pharmacy College of Pharmacy Hanyang University Ansan 15588 Republic of KoreaInstitute of Pharmacological Research Hanyang University Ansan 15588 Republic of Korea) Bae Jonghyun (Department of Pharmacy College of Pharmacy Hanyang University Ansan 15588 Republic of KoreaInstitute of Pharmacological Research Hanyang University Ansan 15588 Republic of Korea) Seo Eunyoung (Department of Pharmacy College of Pharmacy Hanyang University Ansan 15588 Republic of KoreaInstitute of Pharmacological Research Hanyang University Ansan 15588 Republic of Korea) Kim Dong Wook (Department of Pharmacy College of Pharmacy Hanyang University Ansan 15588 Republic of KoreaInstitute of Pharmacological Research Hanyang University Ansan 15588 Republic of Korea) Kim Eun Jin (Department of Pharmacy College of Pharmacy Hanyang University Ansan 15588 Republic of KoreaInstitute of Pharmacological Research Hanyang University Ansan 15588 Republic of Korea)

저널정보: 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제33권 제6호

발행연도: 2023.6

수록면: 736 - 744 (9page)

DOI: 10.4014/jmb.2302.02014

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The introduction of the toxT-139F allele triggers the expression of TCP (toxin co-regulated pilus) and CT (cholera toxin) under simple laboratory culture conditions in most Vibrio cholerae strains. Such V. cholerae strains, especially strains that have been used in OCVs (oral cholera vaccines), can induce antibody responses against TCP in animal models. However, CT produced in these V. cholerae strains is secreted into the culture medium. In this study, V. cholerae strains that can express intracellular CTB under the control of the toxT-139F allele have been constructed for potential application in OCVs. First, we constructed a recombinant plasmid directly linking the ctxAB promoter to ctxB without ctxA and confirmed CTB expression from the plasmid in V. cholerae containing the toxT-139F allele. We constructed another recombinant plasmid to express NtrCTB, from which 14 internal amino acids—from the 7th to the 20th amino acid—of the leader peptide of CTB have been omitted, and we found that NtrCTB remained in the cells. Based on those results, we constructed V. cholerae strains in which chromosomal ctxAB is replaced by ntrctxB or ntrctxB-dimer. Both NtrCTB and NtrCTB-dimer remained in the bacterial cells, and 60% of the NtrCTB-dimer in the bacterial cells was maintained in a soluble form. To develop improved OCVs, these strains could be tested to see whether they induce immune responses against CTB in animal models.

#Cholera #cholera toxin (CT) #cholera toxin B subunit (CTB) #Vibrio cholerae #vaccine

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