质粒pBR322在产气克雷伯菌NCTC 418中的电穿孔及表达，以及质粒pRK2501在恶臭假单胞菌CYM 318中的电穿孔及表达。

Electroporation and expression of plasmid pBR322 in Klebsiella aerogenes NCTC 418 and plasmid pRK2501 in Pseudomonas putida CYM 318.

作者信息

Trevors J T

机构信息

Department of Environmental Biology, University of Guelph, Ontario, Canada.

出版信息

J Basic Microbiol. 1990;30(1):57-61. doi: 10.1002/jobm.3620300116.

DOI:10.1002/jobm.3620300116

PMID:2187074

Abstract

Klebsiella aerogenes NCTC 418 and Pseudomonas putida CYM 318 were transformed via high-voltage electroporation with plasmids pBR322 and pRK2501, respectively. The number of transformants obtained was dependent on the applied voltage, capacitance, and cell recovery procedure. For example, 7.87 x 10(4) transformants/micrograms DNA were obtained at 2500 V, 25 muF when K. aerogenes cells were electroporated with pBR322 DNA. A lower voltage (1500) and capacitance (3 muF) yielded 2.4 x 10(3) transformants/micrograms DNA. P. putida CYM 318 required a 24 h outgrowth period to assist in the recovery of transformants containing pRK2501. Electroporation may be a useful protocol to transform bacterial strains that are not easily transformed by traditional methods.

摘要

产气克雷伯菌NCTC 418和恶臭假单胞菌CYM 318分别通过高压电穿孔法用质粒pBR322和pRK2501进行转化。获得的转化子数量取决于所施加的电压、电容和细胞复苏程序。例如，当产气克雷伯菌细胞用pBR322 DNA进行电穿孔时，在2500 V、25 μF条件下可获得7.87×10⁴个转化子/微克DNA。较低的电压（1500 V）和电容（3 μF）产生2.4×10³个转化子/微克DNA。恶臭假单胞菌CYM 318需要24小时的生长延滞期来帮助含有pRK2501的转化子复苏。电穿孔可能是一种用于转化不易通过传统方法转化的细菌菌株的有用方法。