Chervatiuk N V, Tovkach F I, Gorb T E
Mikrobiol Z. 2007 Sep-Oct;69(5):35-41.
A method for transformation of Erwinia carotovora subsp. carotovora cells has been developed. Transformants of E. carotovora RC 5297 (pECL 18) have been obtained. The transformation frequency was 5.4 x10(2) per 1 microg of DNA. This value depends considerably on the culture growth stage and the amount of calcium chloride used for obtaining the competent cells. The spontaneous loss of the plasmid is below 0.1% per one generation of cells. Both the plasmid and the host RM-systems behave autonomously, i.e., they restrict the development of phages independent of each other. Results obtained are a precondition for creating vectors based on E. carolovora being nonpathogenic for people, endogenic and exogenic plasmids.
已开发出一种用于胡萝卜软腐欧文氏菌胡萝卜软腐亚种细胞转化的方法。已获得胡萝卜软腐欧文氏菌RC 5297（pECL 18）的转化体。转化频率为每1微克DNA 5.4×10² 。该值在很大程度上取决于培养物的生长阶段以及用于制备感受态细胞的氯化钙用量。质粒的自发丢失率低于每代细胞0.1%。质粒和宿主RM系统均自主发挥作用，即它们相互独立地限制噬菌体的发育。所获得的结果是构建基于对人无致病性的胡萝卜软腐欧文氏菌、内生和外生质粒的载体的前提条件。
