Characterization of Tn10d-Cam: a transposition-defective Tn10 specifying chloramphenicol resistance

Mol Gen Genet. 1988 Aug;213(2-3):332-8. doi: 10.1007/BF00339599.

Abstract

We have constructed a small, transposition-defective derivative of the transposon Tn10 that carries the chloramphenicol acetyltransferase gene of pACYC184. This new genetic element, Tn10d-Cam, transposes when Tn10 transposase is provided from a multi-copy plasmid. Transposon insertion mutagenesis of Salmonella typhimurium was performed by using a strain carrying a Tn10d-Cam insertion in an Escherichia coli F' episome as the donor in transductional crosses into recipients that carried a plasmid expressing Tn10 transposase. Tn10d-Cam insertion mutations were also generated by complementation in cis of Tn10d-Cam by a cotransducible Tn10 element that overproduces transposase. Here, transposase was provided only transiently, and the Tn10d-Cam insertion mutations were recovered in a transposase-free strain. Cis complementation was used for mutagenesis of a plasmid target. The site specificity of insertion and the effect of insertions on expression of a downstream gene were investigated, using Tn10d-Cam insertions in a plasmid carrying a segment of the histidine operon.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Chloramphenicol Resistance / genetics*
  • DNA Transposable Elements*
  • Escherichia coli / genetics
  • Genes, Bacterial
  • Genetic Complementation Test
  • Mutation
  • Plasmids
  • Salmonella typhimurium / genetics

Substances

  • DNA Transposable Elements