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

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

J M Pemberton  1 , R J Penfold
Affiliations

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

J M Pemberton et al. Curr Microbiol. 1992 Jul.

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.

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