通过电穿孔法对铜绿假单胞菌进行转化。

Transformation of Pseudomonas aeruginosa by electroporation.

作者信息

Diver J M, Bryan L E, Sokol P A

机构信息

Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.

出版信息

Anal Biochem. 1990 Aug 15;189(1):75-9. doi: 10.1016/0003-2697(90)90046-c.

DOI:10.1016/0003-2697(90)90046-c

PMID:2126169

Abstract

Optimum conditions were defined for the electrotransformation of Pseudomonas aeruginosa PAO1 with plasmid pLAFR1, resulting in a 1500-fold increase in transformation efficiency compared to conventional chemical transformation with MgCl2. In addition, PAO236 and two out of three recent clinical isolates of P. aeruginosa from the sputum of cystic fibrosis patients were successfully transformed with plasmid pUC19 1.8. The applied voltage and the electroporation buffer composition were shown to have the greatest effect on transformation efficiency. Freezing the cells and prolonged storage at -70 degrees C did not significantly affect the transformation efficiency. The clinical isolates tested had lower transformation efficiencies than PAO1.

摘要

确定了用质粒pLAFR1对铜绿假单胞菌PAO1进行电转化的最佳条件，与用MgCl2进行的传统化学转化相比，转化效率提高了1500倍。此外，PAO236以及来自囊性纤维化患者痰液的三株近期铜绿假单胞菌临床分离株中的两株，成功用质粒pUC19 1.8进行了转化。结果表明，施加的电压和电穿孔缓冲液组成对转化效率影响最大。冷冻细胞并在-70℃下长期保存对转化效率没有显著影响。所测试的临床分离株的转化效率低于PAO1。