Strikhanov S N, Aleshkin G I, Skavronskaia A G
Mol Gen Mikrobiol Virusol. 1985 May(5):13-9.
The restriction endonuclease EcoRI dependent recombination of compatible plasmids has been studied in RecA cells of Escherichia coli. Plasmid RP4 and the isogenic ColE1 type plasmids pSA14 or pSA25, differing in restriction-modification RM EcoRI genes, have been used to study this type of recombination. EcoRI dependent recombination of plasmids is demonstrated in RecA cells and, thus, is independent of general system of homologous recombination. The classes of recombinant plasmids isolated from RecA cells differ from the classes isolated from wild type cells. Levels of tetracycline resistance conferred by plasmid RP4 are shown to be dependent on the alleles of RecA+ gene, being extremely low in RecA cells. This property is demonstrated to be useful for obtaining the multicopy recombinant plasmids resulting from EcoRI dependent recombination in RecA cells of Escherichia coli.
已在大肠杆菌的RecA细胞中研究了依赖限制性内切酶EcoRI的相容性质粒重组。质粒RP4以及在限制性修饰RM EcoRI基因方面存在差异的同基因ColE1型质粒pSA14或pSA25已被用于研究此类重组。在RecA细胞中证实了质粒的EcoRI依赖性重组,因此,它独立于同源重组的一般系统。从RecA细胞中分离出的重组质粒类别与从野生型细胞中分离出的类别不同。结果表明,质粒RP4赋予的四环素抗性水平取决于RecA+基因的等位基因,在RecA细胞中极低。已证明该特性有助于在大肠杆菌的RecA细胞中获得由EcoRI依赖性重组产生的多拷贝重组质粒。