An improved counterselection cassette for use in Haemophilus influenzae

Gene. 2012 Jan 15;492(1):325-8. doi: 10.1016/j.gene.2011.10.014. Epub 2011 Oct 20.

Abstract

Counterselectable cassettes are extremely useful in molecular biology and allow for the creation of unmarked deletion mutants or the introduction of point mutations. I have constructed an inducible sacB cassette, using the tetracycline repressor. When used in tandem with a kanamycin-resistance marker, the cassette was successful in creating unmarked mutants in Haemophilus influenzae. The inducible nature of the cassette avoids some of the common problems associated with the utilization of sacB in counterselection.

MeSH terms

  • Bacillus subtilis / genetics
  • Genes, Bacterial
  • Genetic Markers*
  • Haemophilus influenzae / genetics*
  • Hexosyltransferases / genetics
  • Kanamycin Resistance / genetics
  • Mutagenesis, Insertional*
  • Mutation*

Substances

  • Genetic Markers
  • Hexosyltransferases
  • levansucrase