Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2000 Nov;68(11):6391-7.
doi: 10.1128/IAI.68.11.6391-6397.2000.

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

S L Chiang  1 , J J Mekalanos
Affiliations

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

S L Chiang et al. Infect Immun. 2000 Nov.

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.

PubMed Disclaimer

Figures

FIG. 1
FIG. 1
Construction of pSC138. Abbreviations: Ap, ampicillin resistance; Cm, chloramphenicol resistance; Km, kanamycin resistance; R6K, oriR6K conditional origin of replication; RP4, origin of transfer; tnp, transposase; HA, influenza virus hemagglutinin epitope.
FIG. 2
FIG. 2
Diagram of construction of nonmotile, Ogawa+ strains using TnFC-rfbT.
See this image and copyright information in PMC

References

    1. Aidara A, Koblavi S, Boye C S, Raphenon G, Gassama A, Grimont F, Grimon P A. Phenotypic and genotypic characterization of Vibrio cholerae isolates from a recent cholera outbreak in Senegal: comparison with isolates from Guinea-Bissau. Am J Trop Med Hyg. 1998;58:163–167. - PubMed
    1. Albert M J, Siddique A K, Islam M S, Faruque A S G, Ansaruzzaman M, Faruque S M, Sack R B. Large outbreak of clinical cholera due to Vibrio cholerae non-O1 in Bangladesh. Lancet. 1993;341:704. - PubMed
    1. Ausubel F M, Brent R, Kingston R E, Moore D E, Seidman J G, Smith J A, Struhl K, editors. Current protocols in molecular biology. New York, N.Y: John Wiley & Sons, Inc.; 1995.
    1. Barua D. History of cholera. In: Barua D, Greenough III W B, editors. Cholera. New York, N.Y: Plenum Publishing Corporation; 1992. pp. 1–36.
    1. Bennish M L. Cholera: pathophysiology, clinical features, and treatment. In: Wachsmuth I K, Blake P A, Olsvik Ø, editors. Vibrio cholerae and cholera: molecular to global perspectives. Washington, D.C.: ASM Press; 1994. pp. 229–255.

Publication types

LinkOut - more resources