作　　者： ; ; ; ; ;

机构地区： 西北农林科技大学动物科技学院陕西省农业分子生物学重点实验室

出　　处： 《华南农业大学学报》 2011年第3期92-96,共5页

摘　　要： 利用鸟枪法通过大肠埃希菌-枯草芽孢杆菌启动子探针载体分离到枯草芽孢杆菌168菌株的103个基因启动子片段,发现1个672 bp的启动子片段P3.4.23,能够在大肠埃希菌和枯草芽孢杆菌高效启动报告基因β-半乳糖苷酶的表达,其酶活性分别达到3 418、2 877 U/mL.相似性分析表明P3.4.23启动子片段具有枯草芽孢杆菌基因启动子的保守序列,并确定其转录活性区域位于5～333 bp,为yxiE基因的调控序列.将黄牛GHRL基因导入到启动子亚克隆序列下游,使该基因在枯草芽孢杆菌中得到了表达. Through shot-gun method,one hundred and three DNA segments of promoter from genomic library of 168 Bacillus subtilis were detected with Escherichia coli-B.subtilis shuttle plasmid.The activities of the reporter gene of bgaB driven by one promoter fragment,P3.4.23,exhibited high expression strength both in E.coli and B.subtilis.The fragment was 672 bp long and stronger than P43 promoter.The activities of bgaB promoted by the fragment reached to 3 418 and 2 877 U/mL in E.coli and B.subtilis,respectively.The blasting result showed that the fragment had the conserved sequences of B.subtilis promoters.The putative promoter was sub-cloned and identified,which showed that its important region for transcription,5-333 bp,is the regulatory region of yxiE gene.GHRL gene of cattle was drove by the sub-cloned promoter.The SDS-PAGE showed that GHRL gene was expressed in B.subtilis.

关 键 词： 枯草芽孢杆菌 大肠埃希菌 启动子探针载体 基因 牛 鸟枪法

领　　域： [农业科学] [农业科学] [农业科学]