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MudSacI, a transposon with strong selectable and counterselectable markers: use for rapid mapping of chromosomal

M Lawes1, S Maloy

  • 1Department of Microbiology, University of Illinois, Urbana 61801.

Journal of Bacteriology
|March 1, 1995
PubMed

Insights

Researchers developed MudSacI, a novel mini-Mu transposon. This tool uses the Bacillus subtilis sacB gene to confer sucrose sensitivity, enabling efficient counterselection for genetic mapping in bacteria like Salmonella typhimurium.

Area of Science:

  • Microbiology
  • Bacteriology
  • Molecular Biology

Background:

  • Transposable bacteriophage Mu and its mini-Mu derivatives are valuable for bacterial genetic analysis.
  • Existing Mu derivatives allow direct selection but often lack effective counterselection methods.
  • Counterselection is crucial for streamlining genetic analysis and mapping.

Purpose of the Study:

  • To construct a novel mini-Mu derived transposon for counterselection.
  • To introduce the Bacillus subtilis sacB gene into a mini-Mu transposon.
  • To demonstrate the utility of this transposon for rapid genetic mapping.

Main Methods:

  • Construction of the MudSacI mini-Mu transposon.
  • Inclusion of the Bacillus subtilis sacB gene conferring sucrose sensitivity.
  • Application of MudSacI for genetic mapping in Salmonella typhimurium.

Main Results:

  • Successful construction of MudSacI, a mini-Mu transposon conferring sucrose sensitivity.
  • Demonstration of MudSacI's effectiveness in rapid genetic mapping of chromosomal genes.
  • Identification of sucrose sensitivity as a viable counterselection marker.

Conclusions:

  • MudSacI provides a powerful counterselection tool for bacterial genetics.
  • This transposon facilitates rapid genetic mapping in Salmonella typhimurium.
  • The approach is adaptable for genetic analysis in a wide range of bacteria.

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