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人DNA聚合酶δ催化亚基cDNA的分子克隆

Molecular cloning of the cDNA for the catalytic subunit of human DNA polymerase delta.

作者信息

Yang C L, Chang L S, Zhang P, Hao H, Zhu L, Toomey N L, Lee M Y

机构信息

Department of Medicine, University of Miami School of Medicine, FL 33101.

出版信息

Nucleic Acids Res. 1992 Feb 25;20(4):735-45. doi: 10.1093/nar/20.4.735.

Abstract

The cDNA of human DNA polymerase delta was cloned. The cDNA had a length of 3.5 kb and encoded a protein of 1107 amino acid residues with a calculated molecular mass of 124 kDa. Northern blot analysis showed that the cDNA hybridized to a mRNA of 3.4 kb. Monoclonal and polyclonal antibodies to the C-terminal 20 residues specifically immunoblotted the human pol delta catalytic polypeptide. A multiple sequence alignment was constructed. This showed that human pol delta is closely related to yeast pol delta and the herpes virus DNA polymerases. The levels of pol delta message were found to be induced concomitantly with DNA pol delta activity and DNA synthesis in serum restimulated proliferating IMR90 cultured cells. The human pol delta gene was localized to chromosome 19 by Southern blotting of EcoRI digested DNA from a panel of rodent/human cell hybrids.

摘要

克隆了人DNA聚合酶δ的cDNA。该cDNA长度为3.5 kb,编码一个由1107个氨基酸残基组成的蛋白质,计算分子量为124 kDa。Northern印迹分析表明,该cDNA与一个3.4 kb的mRNA杂交。针对C末端20个残基的单克隆和多克隆抗体特异性免疫印迹人pol δ催化多肽。构建了多序列比对。结果表明,人pol δ与酵母pol δ和疱疹病毒DNA聚合酶密切相关。在血清再刺激的增殖性IMR90培养细胞中,发现pol δ信使水平与DNA pol δ活性和DNA合成同时被诱导。通过对一组啮齿动物/人类细胞杂种的EcoRI消化DNA进行Southern印迹分析,将人pol δ基因定位到19号染色体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2b6/312012/c7fa7cd8840f/nar00078-0096-a.jpg

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