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DNA聚合酶III全酶ψ亚基编码基因的鉴定、分离及过表达

Identification, isolation, and overexpression of the gene encoding the psi subunit of DNA polymerase III holoenzyme.

作者信息

Carter J R, Franden M A, Aebersold R, McHenry C S

机构信息

Department of Biochemistry, Biophysics and Genetics, University of Colorado Health Sciences Center, Denver 80262.

出版信息

J Bacteriol. 1993 Sep;175(17):5604-10. doi: 10.1128/jb.175.17.5604-5610.1993.

Abstract

The gene encoding the psi subunit of DNA polymerase III holoenzyme, holD, was identified and isolated by an approach in which peptide sequence data were used to obtain a DNA hybridization probe. The gene, which maps to 99.3 centisomes, was sequenced and found to be identical to a previously uncharacterized open reading frame that overlaps the 5' end of rimI by 29 bases, contains 411 bp, and is predicted to encode a protein of 15,174 Da. When expressed in a plasmid that also expressed holC, holD directed expression of the psi subunit to about 3% of total soluble protein.

摘要

通过一种利用肽序列数据获得DNA杂交探针的方法,鉴定并分离出了编码DNA聚合酶III全酶psi亚基的基因holD。该基因定位于99.3厘摩,经过测序发现它与一个先前未鉴定的开放阅读框相同,该开放阅读框与rimI的5'端重叠29个碱基,包含411个碱基对,预计编码一个15174道尔顿的蛋白质。当在一个也表达holC的质粒中表达时,holD指导psi亚基的表达量达到总可溶性蛋白的约3%。

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