A new method for integration and stable DNA amplification in poorly transformable bacilli

FEMS Microbiol Lett. 1995 Jan 1;125(1):107-14. doi: 10.1111/j.1574-6968.1995.tb07343.x.

Abstract

We have developed a strategy for the integration and stable amplification of DNA sequences in the chromosome of poorly transformable bacilli, which avoids the presence of a functional plasmid replication system in the integrated DNA. The parental vector for integration contains two plus origins of replication from pUB110 in the same orientation on a single plasmid. Due to the direct repeats, such plasmids produce two individual progeny vectors, one of which is dependent on the other for replication, as it lacks a functional rep gene. We have used such a progeny vector system to integrate and amplify DNA on the chromosome of Bacillus licheniformis, and show that the structure is stable in the absence of selective pressure.

MeSH terms

  • Bacillus / genetics*
  • Bacillus subtilis / genetics
  • Chromosomes, Bacterial
  • DNA, Bacterial / genetics*
  • Escherichia coli / genetics*
  • Genetic Vectors
  • Nucleic Acid Amplification Techniques*
  • Plasmids
  • Restriction Mapping
  • Transformation, Bacterial*

Substances

  • DNA, Bacterial