Cloning of the structural gene (hly) for the haemolysin of Vibrio cholerae El Tor strain 017

Paul A. Manning, Melissa H. Brown, Michael W. Heuzenroeder

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38 Citations (Scopus)

Abstract

We have cloned the DNA encoding the haemolysin of Vibrio cholerae El Tor strain 017 into the plasmid vector pBR322. The resultant plasmid, pPM431, has a 6.2-kb PstI DNA insert which leads to the production of the haemolysin in Escherichia coli K-12. Deletion analysis and transposon mutagenesis have allowed us to localize several regions affecting haemolysin production. A number of these mutants have been analysed in E. coli K-12 minicells. Three proteins have been identified: A, 80 kDal; B, 71 kDal; and C, 22 kDal. A is the haemolysin which appears to be cell-associated in E. coli K-12, and B and C are required for its efficient production. We suggest that the genes for proteins A, B and C be designated hlyA, hlyB and hlyC, respectively.

Original languageEnglish
Pages (from-to)225-231
Number of pages7
JournalGene
Volume31
Issue number1-3
DOIs
Publication statusPublished - Nov 1984
Externally publishedYes

Keywords

  • E. coli K-12
  • minicells
  • physical map
  • protein secretion
  • Recombinant DNA
  • transposon mutagenesis

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