AUTONOMOUS REPLICATION OF PLASMIDS CONTAINING DNA FRAGMENTS FROM THE ALPHA-POLYMERASE COMPLEX OF RAT-LIVER

作者：TIMCHENKO NA^*; SHCHERBAKOV OG; KRUTYAKOV VM; FILATOV MV

来源：Molecular Biology, 1990, 24(3): 673-680.

A group of plasmids designated pori, containing DNA fragments from the rat liver alpha-polymerase complex was prepared. Colony hybridization yielded 30 clones homologous to the SV40 virus replicatin initiation region. Five pori plasmids were transfected in Chinese hamster V79 fibroblasts. Plasmid DNA was extracted at varous times after transfection, and resistance to the restriction enzyme CfuI and incorporation of bromodeoxyuridine into plasmid DNA were determined. Two plasmids, pori32 and pori22, were able to replicate autonomously in mammalian cells. Thus, at least two DNA fragments from the alpha-polymerase complex function as replication initiation regions in mammalian cells. Vector (pUC19) transcrips were detected five days after transfection of V79 cells with pori32. The results show that the pori32 sequence contains replication initiation and promoter sites.

• 出版日期1990-6