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Serotype conversion in Vibrio cholerae O1.

U H Stroeher1, L E Karageorgos, R Morona

  • 1Department of Microbiology and Immunology, The University of Adelaide, South Australia.

Proceedings of the National Academy of Sciences of the United States of America
|April 1, 1992
PubMed
Summary

The Vibrio cholerae O1 serotype conversion from Inaba to Ogawa is controlled by the rfbT gene. This gene

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Vibrio cholerae O1 comprises Inaba and Ogawa serotypes, distinguished by their O antigen lipopolysaccharide.
  • These serotypes can interconvert, a phenomenon with implications for cholera outbreaks.
  • Understanding the genetic basis of this interconversion is crucial for disease control.

Purpose of the Study:

  • To elucidate the genetic mechanisms underlying Vibrio cholerae O1 serotype conversion.
  • To identify the specific gene responsible for the Inaba to Ogawa serotype switch.
  • To characterize the function and origin of the identified gene.

Main Methods:

  • Cloning and sequencing of the 20-kilobase rfb regions involved in O-antigen biosynthesis.
  • Site-directed mutagenesis of the rfbT gene in the Ogawa strain O17.
  • Functional complementation assays using the rfbT gene.
  • Southern hybridization and PCR sequencing of various Vibrio cholerae strains.
  • Protein expression and identification in Escherichia coli K-12.
  • Primer extension studies to define mRNA 5' end.

Main Results:

  • A single nucleotide change creating a stop codon in the rfbT gene was identified in the Inaba strain 569B.
  • The rfbT gene was confirmed as essential for the Inaba to Ogawa serotype conversion.
  • Mutating rfbT in the Ogawa strain prevented conversion, while complementation restored it.
  • Observed genetic changes in rfbT were not conserved across all Inaba strains, explaining variable conversion.
  • The rfbT gene encodes a 32-kDa protein, transcribed separately, with a G+C content suggesting non-Vibrio origin.

Conclusions:

  • The rfbT gene is the key determinant for the Ogawa serotype in Vibrio cholerae O1.
  • Variations in rfbT sequence and expression influence the ability of Inaba strains to convert to Ogawa.
  • The rfbT gene's likely horizontal acquisition indicates a dynamic evolutionary history for Vibrio cholerae O1.

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