光合菌Rhodobacter sphaeroides 601 hupT基因的克隆与功能分析  

Sequence and Function Analysis of hupT Gene in Rhodobacter sphaeroides 601

作　　者：徐冬青[1] 吴永强[1] 

机构地区：[1]中国科学院上海植物生理研究所,上海200032

出　　处：《植物生理学报（0257-4829)》2001年第6期509-514,共6页Acta Phytophysiologica Sinica

基　　金：国家自然科学基金资助项目 (编号 39870 0 0 5 )~~

摘　　要：从紫色非硫光合细菌Rhodobactersphaeroides60 1的吸氢酶 (hup)基因簇中 ,克隆了hupT基因 ,并对该基因进行了测序 ,分析了由其推测的氨基酸序列的同源性。hupT基因全长 13 3 2bp ,编码一分子量约为 4 8.2 3kD的蛋白。将hupT基因引入大肠杆菌进行了体外表达。纯化基因产物HupT ,并进行HupT的自身磷酸化分析。结果表明 ,HupT属于双组份调节系统中的组氨酸蛋白激酶。将hupT基因导入光合菌Rhodobactercapsulatus吸氢酶负调节基因突变株BSE8后 ,野生型吸氢酶的表型得以恢复 ,说明所克隆的R .sphaeroides 60The hupT gene from hup cluster(cosmid I) in Rhodobacter sphaeroides was cloned and sequenced. It has been found to have a length of 1 332 bp and to encode a 48.23 kD protein homologous to histidine kinase belonging to the superfamily of two component regulatory system(Figs.1,2). The HupT was overexpressed in E.coli and purified(Fig.3). The auto phosphorylation of HupT showed that it was a histidine kinase(Fig.4). And the hupT gene could restore hydrogenase activity in R.capsulatus HupT mutant(Table 2).

关 键 词：光合细菌 浑球红细菌 吸氢酶 hupT基因 组氨酸蛋白激酶 基因克隆 

分 类 号：Q785[生物学—分子生物学]