화학공학소재연구정보센터
Current Microbiology, Vol.25, No.1, 25-29, 1992
HIGH-FREQUENCY ELECTROPORATION AND MAINTENANCE OF PUC-BASED AND PBR-BASED CLONING VECTORS IN PSEUDOMONAS-STUTZERI
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/mu-g 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.