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

Autor: Pemberton JM; Microbiology Department, University of Queensland, St. Lucia, Brisbane, Australia., Penfold RJ
Jazyk: angličtina
Zdroj: Current microbiology [Curr Microbiol] 1992 Jul; Vol. 25 (1), pp. 25-9.
DOI: 10.1007/BF01570078
Abstrakt: 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.
Databáze: MEDLINE