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嗜盐古菌沃氏嗜盐碱杆菌的二氢硫辛酰胺脱氢酶：克隆基因的同源过表达

Dihydrolipoamide dehydrogenase from the halophilic archaeon Haloferax volcanii: homologous overexpression of the cloned gene.

作者信息

Jolley K A, Rapaport E, Hough D W, Danson M J, Woods W G, Dyall-Smith M L

机构信息

School of Biology and Biochemistry, University of Bath, United Kingdom.

出版信息

J Bacteriol. 1996 Jun;178(11):3044-8. doi: 10.1128/jb.178.11.3044-3048.1996.

DOI:10.1128/jb.178.11.3044-3048.1996
PMID:8655478
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC178050/
Abstract

The gene encoding dihydrolipoamide dehydrogenase from the halophilic archaeon, Haloferax volcanii, has been subcloned and overexpressed in the parent organism by using the halophilic archaeal rRNA promoter. The recombinant protein has been purified to homogeneity and characterized with respect to its kinetic, molecular, and salt-dependent properties. A dihydrolipoamide dehydrogenase-minus mutant of H. volcanii has been created by homologous recombination with the subcloned gene after insertion of the mevinolin resistance determinant into the protein-coding region. To explore the physiological function of the dihydrolipoamide dehydrogenase, the growth properties of the mutant halophile have been examined.

摘要

来自嗜盐古菌沃氏嗜盐栖热菌（Haloferax volcanii）的二氢硫辛酰胺脱氢酶编码基因已被亚克隆，并通过使用嗜盐古菌rRNA启动子在亲本生物体中过表达。重组蛋白已被纯化至同质，并对其动力学、分子和盐依赖性特性进行了表征。通过将美伐他汀抗性决定簇插入蛋白质编码区后，利用亚克隆基因进行同源重组，构建了沃氏嗜盐栖热菌的二氢硫辛酰胺脱氢酶缺失突变体。为了探索二氢硫辛酰胺脱氢酶的生理功能，已对突变嗜盐菌的生长特性进行了研究。

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