中度嗜盐菌Halomonas sp. BYS-1启动子的克隆和测序  

CLONING AND SEQUENCING OF PROMOTERS OF A MODERATE HALOPHILIC BACTERIUM HALOMONAS SP. BYS-1

作　　者：洪青[1] 张忠辉[1] 张晓舟[1] 徐剑宏[1] 李顺鹏[1] 

机构地区：[1]南京农业大学生命科学学院农业部农业环境微生物工程重点开放实验室

出　　处：《应用与环境生物学报》2005年第6期729-732,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然基金项目(No.30400013);南京农业大学青年科技创新项目(No.KJ04018)资助~~

摘　　要：提取了中度嗜盐菌Halomonassp.BYS1的基因组DNA,以甲基对硫磷水解酶基因(mpd)为报告基因,以启动子探针pUCmpd为载体,通过鸟枪法在E.coliDH5α中构建了BYS1的启动子文库.通过筛选获得了17个阳性克隆,编号为P1～P17.测定了阳性克隆的甲基对硫磷水解酶(MPH)活性,结果表明,P3中mpd基因的启动子活性最强,它的酶活高达2554.3U/mg,而P17中mpd基因的启动子活性最弱,它的酶活只有68.3U/mg.对P3、P8、P17克隆中的重组质粒的插入片段进行了测序和在线启动子预测.High quality genomic DNA of a moderately halophilic bacterial strain Halomonas sp. BYS-1 was extracted. The promoter library of BYS-1 was constructed in E. coli DH5α with pUC-mpd as the promoter probe vector by shotgun-cloning method. 17 positive clones designated as P1 -P17 were obtained by screening. Their methyl parathion hydrolase （MPH） activity was assayed to compare the promoting strength of the promoters.The results showed that the promoter of mpd in P3 was the strongest with MPH activity of 2 554.3 U/mg,while the promoter of mpd in P17 was the weakest of 68.3 U/mg. The inserting fragments in the recombinant plasmids from P3, P8 and P17 were sequenced and subjected to online promoter prediction.Fig 4, Tab 1, Ref 17

关 键 词：中度嗜盐菌 启动子 克隆 测序 

分 类 号：Q78[生物学—分子生物学]