少根根霉脂肪酶基因在枯草芽孢杆菌中的诱导表达  

Inducible Expression of Rhizopus arrhizus Lipase Gene in Bacillus subtilis

作　　者：张欢[1] 王凤寰[1] 田平芳[1] 谭天伟[1] 李文进[1] 

机构地区：[1]北京化工大学生命科学与技术学院北京市生物加工过程重点实验室,北京100029

出　　处：《生物技术通报》2010年第1期123-127,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(20876011);国家"863"计划项目(2006AA020203);国家"973"计划项目(2007CB707804);北京市自然基金项目(2071002);北京市教育委员会共建项目;生物炼制教育部工程研究中心资助项目

摘　　要：利用PCR技术从少根根霉基因组中扩增出脂肪酶成熟肽基因ral,并从枯草芽孢杆菌基因组中扩增出sacB基因的启动子-信号序列(SacB);通过搭桥PCR将SacB序列与ral基因融合,并将该基因表达盒连接到枯草杆菌分泌表达载体pGJ103中构建了脂肪酶基因的诱导表达载体pGJ103-SacB-ral。将重组载体转化至枯草芽孢杆菌后,少根根霉脂肪酶成熟肽基因在SacB启动子-信号序列的调控和蔗糖的诱导下获得表达,产物分泌至胞外。In this research,lipase mature peptide general and a promoter-signal fragment （SacB）of gene sacB from genomic DNA of Rhizopus arrhizus and Bacillus subtili ,was cloned respectively, by PCR. To obtain efficient expression of gene ral, strategy of pGJ103 fusion expression was employed between SacB fragment and general by overlap-PCR. Thus,an inducible recombinant plasmid pGJ103- SacB-ral containing the fusion expression cassette was constructed based on the secreted expression plasmid pGJ103 of B. subtili. After the competent ceils of B. subtilis were transformed with the recombinant plasmid,upon regulation of SacB fragment and inducement of saccharose,gene ral was able to be expressed and its production was examined from the outside of the cells.

关 键 词：少根根霉 脂肪酶成熟肽 枯草芽孢杆菌 SacB 诱导表达 

分 类 号：Q78[生物学—分子生物学]