Use of recombinase gene fusions to identify Vibrio cholerae genes induced during infection.

Abstract:

:A complete understanding of host-parasite interactions must necessarily include the identification and characterization of gene products expressed by both parties during the infectious process. We have developed a new screen to identify bacterial genes that are transcriptionally induced during infection of a host animal. The method is based on pre-selection of strains carrying tnpR operon fusions (encoding resolvase, a site-specific DNA recombinase) which are not expressed in vitro, followed by screening for a subset of these strains that subsequently express resolvase within the host environment. The latter subset was recognized as recombinants that had deleted a resolvase-specific reporter construct. Thirteen transcription units of Vibrio cholerae were identified that were induced during infection in an infant mouse model of cholera. Five of these were predicted to encode polypeptides with diverse functions in metabolism, biosynthesis and motility; one encoded a secreted lipase; two appear to be antisense to genes involved in motility; and five are predicted to encode polypeptides of unknown function. Three of the transcripts were shown to be required for full virulence in infant mice, as assessed by competition experiments.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Camilli A,Mekalanos JJ

doi

10.1111/j.1365-2958.1995.mmi_18040671.x

subject

Has Abstract

pub_date

1995-11-01 00:00:00

pages

671-83

issue

4

eissn

0950-382X

issn

1365-2958

journal_volume

18

pub_type

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