枯草芽孢杆菌启动子序列在大肠杆菌中的克隆及序列分析  

Cloning and Sequence Analysis of Promoter Sequence of Bacillus subtilis

作　　者：陈晓月[1] 金宁一[2] 邹伟[1] 海洋[1] 马海利[2] 李昌[2] 

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110161 [2]军事医学科学院军事兽医研究所

出　　处：《沈阳农业大学学报》2008年第3期371-373,共3页Journal of Shenyang Agricultural University

基　　金：辽宁省教育厅科学研究项目(20060786)

摘　　要：根据GenBank中枯草芽孢杆菌的P43启动子基因序列,设计合成了一对引物,用PCR方法钓取枯草芽孢杆菌AS.1655菌株的启动子基因,PCR产物经琼脂糖凝胶回收纯化后,连接到pMD18T-simple载体上,进行序列测定。测序结果表明:枯草芽孢杆菌AS.1655菌株的启动子全长427 bp,由两个叠加的启动元件组成,分别为5σ5和3σ7 RNA聚合酶识别的位点,与GenBank中的其他序列比较发现核苷酸的同源性为99.5%,氨基酸的同源性为98.6%,说明枯草杆菌P43启动子的基因序列高度保守。According to the p^43 promoter gene of bacillus subtilis reported by GenBank, a pair of specific primers were designed and used to amplify promoter gene of AS. 1655 strain. The PCR products were purified and inserted into pMD18-T simple vector. The recombinant plasmid was used to be sequenced. The gene of p^43 was composed with two overlap promoters which could be recognized by σ^55 and σ^37 RNA polymerase respectively. The length of objective gene was 427bp. The homology of nucleic acid and amino acid were 99.5 % and 98.6 % respectively. Compared with other gene in GenBank, the results showed that the p^43 gene was highly conservative.

关 键 词：枯草芽孢杆菌 p^43启动子 克隆 序列分析 

分 类 号：Q782[生物学—分子生物学]