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LexA,一种转录调节因子,可结合在集胞藻PCC 6803双向氢化酶的启动子区域。

LexA, a transcription regulator binding in the promoter region of the bidirectional hydrogenase in the cyanobacterium Synechocystis sp. PCC 6803.

作者信息

Oliveira Paulo, Lindblad Peter

机构信息

Department of Physiological Botany, EBC, Uppsala University, Villavägen 6, SE-752 36 Uppsala, Sweden.

出版信息

FEMS Microbiol Lett. 2005 Oct 1;251(1):59-66. doi: 10.1016/j.femsle.2005.07.024.

Abstract

The unicellular cyanobacterium Synechocystis PCC 6803 contains a single pentameric bidirectional hydrogenase encoded by hoxEFUYH. Transcriptional experiments demonstrated that the five hox genes are part of a single transcript together with three ORFs with unknown functions. The transcription start point was localized by 5' RACE to 168bp upstream the hoxE ATG start codon. DNA affinity assays demonstrated a specific interaction between the hox regulatory promoter region and a protein which, using mass spectrometry, was identified to be LexA. Overexpressed His-tagged Synechocystis LexA and EMSA showed a specific binding to the promoter region of the hox operon. Increasing concentrations of the purified LexA resulted in two retarded LexA-DNA complexes, in agreement with the presence of two putative LexA binding sites upstream the determined TSP.

摘要

单细胞蓝藻集胞藻PCC 6803含有一种由hoxEFUYH编码的单一五聚体双向氢化酶。转录实验表明,五个hox基因与三个功能未知的开放阅读框(ORF)共同构成了一个单一转录本的一部分。转录起始点通过5' RACE定位到hoxE起始密码子ATG上游168bp处。DNA亲和分析表明,hox调控启动子区域与一种蛋白质之间存在特异性相互作用,通过质谱鉴定该蛋白质为LexA。过表达的His标签集胞藻LexA和电泳迁移率变动分析(EMSA)表明,其与hox操纵子的启动子区域存在特异性结合。纯化的LexA浓度增加导致出现两种滞后的LexA-DNA复合物,这与在确定的转录起始点(TSP)上游存在两个假定的LexA结合位点一致。

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