在鱼腥藻PCC 7120中，ManR特异性结合Nramp转运蛋白基因的启动子：蓝细菌中的一种新型调控DNA基序。
The ManR specifically binds to the promoter of a Nramp transporter gene in Anabaena sp. PCC 7120: a novel regulatory DNA motif in cyanobacteria.
作者信息
Huang Wei, Wu Qingyu
机构信息
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, PR China.
出版信息
Biochem Biophys Res Commun. 2004 Apr 30;317(2):578-85. doi: 10.1016/j.bbrc.2004.03.089.
The Anabaena sp. PCC 7120 ManR and a homologous protein of MntH were identified by BLAST search. Recombinant ManR protein was overexpressed in Escherichia coli and purified by an immobilized metal (Ni) affinity chromatography. Electrophoretic mobility shift assays revealed that ManR specifically bound to the promoter region of the mntH gene. Site-directed mutagenesis experiments demonstrated that the specific recognition site for ManR is TATGAAAAGAATATGAGAA, which is composed of two direct repeats of the consensus sequence (T/A)ATGA(G/A)A(A/G). This is a novel regulatory DNA motif in cyanobacteria, indicating that the expression of mntH was regulated by a two-component Mn(2+)-Sensing System containing ManR in Anabaena sp. PCC 7120. To date, this specific pathway of regulating mntH expression has only been found in cyanobacteria.
通过BLAST搜索鉴定出鱼腥藻属PCC 7120的ManR以及MntH的同源蛋白。重组ManR蛋白在大肠杆菌中过量表达，并通过固定化金属（镍）亲和层析进行纯化。电泳迁移率变动分析表明，ManR特异性结合mntH基因的启动子区域。定点诱变实验证明，ManR的特异性识别位点为TATGAAAAGAATATGAGAA，它由共有序列（T/A）ATGA（G/A）A（A/G）的两个直接重复组成。这是蓝细菌中一种新的调控DNA基序，表明在鱼腥藻属PCC 7120中，mntH的表达受包含ManR的双组分锰（2+）传感系统调控。迄今为止，这种调控mntH表达的特定途径仅在蓝细菌中被发现。
Zotero 插件