Aleshkin G I, Strikhanov S N, Skavronskaia A G
Mol Gen Mikrobiol Virusol. 1985 Apr(4):15-21.
The possible participation of restriction endonuclease EcoRI in recombination of compatible nonhomologous plasmids in E. coli cells has been studied. To study the process, plasmids RP4 and R245 have been transferred by conjugation into the recipient cells of E. coli harbouring one of isogenic plasmids, pSA14 and pSA25, different for the genes coding restriction endonuclease EcoRI. The genetic analysis of transconjugant phenotypes, coded by the plasmids, has permitted to register the recombinant plasmids after compatibility of parent plasmids in E. coli cells. Recombination of plasmid RP4 with the plasmid pSA14, carrying EcoRI genes, has been registered in E. coli cells, producing the restriction endonuclease, while plasmid recombination has not been found in the cells harbouring plasmid pSA25, isogenic for all genes, except for EcoRI genes, with plasmid pSA14. Restriction endonuclease EcoRI is concluded to stimulate site specific recombination of nonhomologous compatible plasmids in vivo. EcoRI-mediated recombination of plasmid R245 with plasmid pSA14 is discussed.
研究了限制性内切酶EcoRI在大肠杆菌细胞中兼容非同源质粒重组过程中的可能参与情况。为了研究该过程,通过接合作用将质粒RP4和R245转移到携带等基因质粒pSA14和pSA25之一的大肠杆菌受体细胞中,这两种等基因质粒在编码限制性内切酶EcoRI的基因上存在差异。对由质粒编码的转接合子表型进行遗传分析,使得在大肠杆菌细胞中亲本质粒兼容后能够鉴定出重组质粒。在产生限制性内切酶的大肠杆菌细胞中,已鉴定出质粒RP4与携带EcoRI基因的质粒pSA14发生了重组,而在除EcoRI基因外所有基因均与质粒pSA14等基因的携带质粒pSA25的细胞中未发现质粒重组。由此得出结论,限制性内切酶EcoRI在体内刺激非同源兼容质粒的位点特异性重组。文中还讨论了EcoRI介导的质粒R245与质粒pSA14的重组。