Transposon-mediated mutagenesis and recombination in Vibrio cholerae

Abstract

An efficient method for introducing transposons into the Vibrio cholerae chromosome or the P plasmid was developed by using F'(ts)lac+ plasmids from Escherichia coli as suicide delivery vehicles. Hybrid P::Tn1, Tn5 and P::Tn1, Tn10 plasmids containing Tn5 or Tn10 in either of the two possible orientations were constructed. During conjugation, these hybrid plasmids mediated oriented transfer of markers from the sites of insertion of homologous transposons in the donor chromosome. The collection of donor strains described here permits oriented mobilization of all parts of the V. cholerae chromosome.