, Volume 25, Issue 1, pp 25-29

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

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Abstract

A number ofEscherichia coli cloning vectors, based on ColE1-like replicons, were shown to be maintained inPseudomonas stutzeri ATCC 17588. A restrictionless mutant ofP. stutzeri was isolated, and this strain was used to develop an efficient electroporation system. With theE. coli cloning vector pHSG298, transformation frequencies of up to 2×107 transformants/μg DNA were achieved. This frequency is comparable to that obtained for CaCl2-mediated transformation ofE. coli; thus, direct cloning of DNA intoP. stutzeri is feasible. As will be discussed, this may prove useful for cloning DNA from high mol% G+C genera in cases in whichE. coli is not a suitable heterologous cloning host.