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编码铁氧化还原蛋白的糖多孢红霉菌fdxA基因的克隆与特性分析

Cloning and characterization of the Saccharopolyspora erythraea fdxA gene encoding ferredoxin.

作者信息

Donadio S, Hutchinson C R

机构信息

Corporate Molecular Biology, Abbott Laboratories, Abbott Park, IL 60064.

出版信息

Gene. 1991 Apr;100:231-5. doi: 10.1016/0378-1119(91)90372-i.

Abstract

The Saccharopolyspora erythraea gene (fdxA) corresponding to a previously purified ferredoxin [Shafiee and Hutchinson, J. Bacteriol., 170 (1988) 1548-1553] was cloned using an oligodeoxyribonucleotide probe based on the N-terminal sequence of the ferredoxin. The nucleotide sequence of a 1.3-kb segment encompassing fdxA indicates that the corresponding protein, SeFdI, is 105 amino acids long, and very similar to other 7Fe ferredoxins. A partial open reading frame closely linked to fdxA was also detected. Disruption of fdxA was attempted by replacing the wild-type allele with an in vitro mutated copy. The failure to construct an fdxA mutant strain suggests that fdxA lies in an essential region of the S. erythraea chromosome.

摘要

对应于先前纯化的铁氧化还原蛋白的糖多孢红霉菌基因(fdxA)[沙菲和哈钦森,《细菌学杂志》,170(1988)1548 - 1553],使用基于该铁氧化还原蛋白N端序列的寡脱氧核糖核苷酸探针进行克隆。包含fdxA的1.3 kb片段的核苷酸序列表明,相应的蛋白质SeFdI长105个氨基酸,并且与其他7Fe铁氧化还原蛋白非常相似。还检测到一个与fdxA紧密相连的部分开放阅读框。试图通过用体外突变的拷贝替换野生型等位基因来破坏fdxA。未能构建出fdxA突变菌株表明fdxA位于糖多孢红霉菌染色体的一个必需区域。

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