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Hfr formation directed by tn10.

F G Chumley1, R Menzel, J R Roth

  • 1Department of Biology, University of Utah, Salt Lake City, Utah 84112.

Genetics
|April 1, 1979
PubMed
Summary
This summary is machine-generated.

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Transposable element Tn10 facilitates the creation of Hfr strains in Salmonella typhimurium by enabling homologous recombination between plasmid and chromosomal Tn10 insertions. This method allows for the isolation of Hfrs with defined origins and transfer directions for genetic mapping.

Area of Science:

  • Microbiology
  • Bacterial Genetics
  • Molecular Biology

Background:

  • Transposable elements like Tn10 are mobile DNA sequences that can insert into various locations within a bacterial genome.
  • F plasmids are conjugative plasmids that can mediate the transfer of genetic material between bacteria, and can also integrate into the chromosome to form Hfr strains.
  • Hfr strains are crucial for bacterial genetic mapping due to their ability to transfer chromosomal DNA in a linear, ordered fashion.

Purpose of the Study:

  • To develop a novel method for generating Hfr strains in Salmonella typhimurium using the transposable element Tn10.
  • To investigate the role of homologous recombination between Tn10 elements in Hfr formation.
  • To establish a system for isolating Hfr strains with predetermined chromosomal origins and transfer directions.

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Main Methods:

  • Construction of F'ts114 lac plasmids carrying Tn10 insertions.
  • Transfer of these engineered plasmids to Salmonella typhimurium strains with chromosomal Tn10 insertions.
  • Selection and characterization of resulting Hfr strains based on their transfer direction and stability.
  • Utilizing recA-dependent homologous recombination between plasmid and chromosomal Tn10 elements.

Main Results:

  • Hfr formation was dependent on the presence of both plasmid and chromosomal Tn10 elements and recA function.
  • All isolated Hfrs from a given merodiploid exhibited the same direction of chromosome transfer.
  • Hfr strains with either forward or reverse transfer directions could be generated by altering the Tn10 orientation in the F' plasmid.
  • The method allowed for the isolation of Hfrs with origins at almost any predetermined chromosomal site, facilitating stable genetic mapping.

Conclusions:

  • The Tn10 element provides a versatile tool for directed Hfr formation in Salmonella typhimurium.
  • This approach enables the precise control over Hfr origin and transfer direction, significantly advancing bacterial genetic mapping strategies.
  • The methodology is broadly applicable to other transposable elements and bacterial species, offering a generalizable technique for genetic analysis.