Construction of a Vibrio cholerae vaccine candidate using transposon delivery and FLP recombinase-mediated excision.

Abstract:

:Recent efforts to develop a vaccine against the diarrheal disease cholera have focused on the use of live attenuated strains of the causative organism, Vibrio cholerae. The Ogawa lipopolysaccharide phenotype is expressed by many epidemic strains, and motility defects reduce the risk of reactive diarrhea in vaccine recipients. We therefore converted a motile Inaba(+) vaccine candidate, Peru-2, to a nonmotile Ogawa(+) phenotype using a mariner-based transposon carrying rfbT, the gene required for expression of the Ogawa phenotype. Analysis of 22 nonmotile Peru-2 mutants showed that two were Ogawa(+), and both of these strains had insertions in the flgE gene. It was possible to convert these strains to antibiotic sensitivity by introducing a recombinase that acts on sites flanking the antibiotic marker on the transposon. The resulting strains are competent for colonization in infant mice and may therefore be suitable as vaccine candidates for use either independently or in a combination with strains of different biotypes and serotypes.

journal_name

Infect Immun

journal_title

Infection and immunity

authors

Chiang SL,Mekalanos JJ

doi

10.1128/iai.68.11.6391-6397.2000

keywords:

subject

Has Abstract

pub_date

2000-11-01 00:00:00

pages

6391-7

issue

11

eissn

0019-9567

issn

1098-5522

journal_volume

68

pub_type

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