High-frequency electroporation and maintenance of pUC- and pBR-based cloning vectors in Pseudomonas stutzeri

Curr Microbiol. 1992 Jul;25(1):25-9. doi: 10.1007/BF01570078.

Abstract

A number of Escherichia coli cloning vectors, based on ColE1-like replicons, were shown to be maintained in Pseudomonas stutzeri ATCC 17588. A restrictionless mutant of P. stutzeri was isolated, and this strain was used to develop an efficient electroporation system. With the E. coli cloning vector pHSG298, transformation frequencies of up to 2 x 10(7) transformants/micrograms DNA were achieved. This frequency is comparable to that obtained for CaCl2-mediated transformation of E. coli; thus, direct cloning of DNA into P. stutzeri is feasible. As will be discussed, this may prove useful for cloning DNA from high mol% G + C genera in cases in which E. coli is not a suitable heterologous cloning host.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriocin Plasmids*
  • Bacteriological Techniques*
  • Carotenoids / biosynthesis
  • Cloning, Molecular / methods*
  • DNA, Bacterial / genetics*
  • Electrochemistry*
  • Genetic Vectors*
  • Pseudomonas / genetics*
  • Replicon
  • Transformation, Bacterial*

Substances

  • DNA, Bacterial
  • Carotenoids