New Gentamicin-Resistance and lacZ Promoter-Probe Cassettes Suitable for Insertion Mutagenesis and Generation of Transcriptional Fusions

Gene. 1995 Aug 30;162(1):37-9. doi: 10.1016/0378-1119(95)00313-u.

Abstract

A set of antibiotic-resistance and promoter-probe cassettes suitable for insertion mutagenesis and generation of transcriptional fusions was constructed. The cassettes contain the aacC1 gene of transposon Tn1696 conferring resistance to gentamicin in a large variety of Gram- and Gram+ bacteria. In addition to the antibiotic-resistance gene, a promoterless Escherichia coli lacZ gene was included in the cassettes, allowing the determination of the transcriptional activity at the insertion site. The cassettes can be excised from a plasmid mediating ampicillin resistance by many commonly used restriction enzymes. The new constructs have been successfully used for mutagenesis and studies of gene transcription in Rhizobium meliloti.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / genetics
  • Base Sequence
  • Drug Resistance, Microbial / genetics
  • Genetic Markers
  • Gentamicins / pharmacology
  • Lac Operon
  • Molecular Sequence Data
  • Mutagenesis, Insertional / methods*
  • RNA / biosynthesis*
  • Selection, Genetic*
  • Transcription, Genetic*

Substances

  • Genetic Markers
  • Gentamicins
  • RNA, recombinant
  • RNA
  • Acetyltransferases
  • gentamicin 3-acetyltransferase