摘要
通过一对自行设计的引物,以E.coli染色体DNA为模板进行PCR扩增,获得完整的recA基因。将PCR产物和pUC18在体外进行连接后,并分别转入E.coliDH5α和E.colik12(λ+),筛选出含重组质粒(pUR4)的转化子。在诱导条件和非诱导条件下,分别测定recA基因在不同转化子中的生物学功能。结果表明recA基因在溶原菌中表现出明显的生理功能,能使λ原噬菌体从溶原状态进入裂解循环。这种重组质粒将在进一步研究λ原噬菌体的诱导机理以及受紫外辐射损伤细胞的修复作用等方面成为有效的工具。
With a pair of primers and the E. coli chromosome DNA as the template, an intact recA gene was obtained by PCR. The recombinant plasmid was constructed by ligating the PCR product and pUC18 in vitro and then transformed into E. coli DH5α and E. coli k12(λ+), respectively. The plasmid pUR4 containing the recA gene has been screened. The biological function of recA gene in different strains was determined with or without UV induction. The results indicated that the recA gene showed a remarkable physiological function in the λ lysogenic bacteria. It could induce the λ prophage to enter the lysis cycle from lysogenic state. This recombinant plasmid will be a useful tool in the studies of the inductive mechanism of λ prophage and the repair of cells damaged by UV-irradiation.
出处
《中国病毒学》
CSCD
2004年第4期394-397,共4页
Virologica Sinica
