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嗜热栖热菌中编码δ1-吡咯啉-5-羧酸还原酶的proC基因的分子克隆及序列分析

Molecular cloning and sequence analysis of the proC gene encoding delta 1-pyrroline-5-carboxylate reductase from an extremely thermophilic eubacterium Thermus thermophilus.

作者信息

Hoshino T, Kosuge T, Hidaka Y, Tabata K, Nakahara T

机构信息

Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.

出版信息

Biochem Biophys Res Commun. 1994 Feb 28;199(1):410-7. doi: 10.1006/bbrc.1994.1244.

Abstract

A gene library of the extremely thermophilic bacterium, Thermus thermophilus HB27, was constructed in Escherichia coli, and recombinant plasmids able to complement proC mutants of HB27 were obtained. Using the plasmids, the complete nucleotide sequence of the proC gene encoding delta 1-pyrroline-5-carboxylate reductase (P5CR) [EC 1.5.1.2] was determined. The deduced amino acid sequence showed a significant homology to those of P5CRs of E. coli, soybean and human. The proC gene of T. thermophilus was expressed in E. coli and the activity of heat-resistant P5CR was determined. The mutation sites of two HB27 proC mutants were also determined.

摘要

构建了嗜热栖热菌HB27在大肠杆菌中的基因文库,并获得了能够互补HB27 proC突变体的重组质粒。利用这些质粒,测定了编码δ1-吡咯啉-5-羧酸还原酶(P5CR)[EC 1.5.1.2]的proC基因的完整核苷酸序列。推导的氨基酸序列与大肠杆菌、大豆和人类的P5CR具有显著同源性。嗜热栖热菌的proC基因在大肠杆菌中表达,并测定了耐热P5CR的活性。还确定了两个HB27 proC突变体的突变位点。

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