枯草芽孢杆菌碱性蛋白酶基因的克隆和表达

Cloning and Expression of Alkaline Protease Gene by Bacillus subtilis

目的:获得碱性蛋白酶基因。方法:用PCR的方法从枯草芽孢杆菌A-109中扩增碱性蛋白酶基因(apr),并进行测序分析,构建表达载体,最后转化大肠杆菌BL21,SDS-聚丙烯酰胺凝胶电泳检测该基因的表达情况。结果:apr基因片段含1092个碱基对。该基因片段核苷酸序列与Bacillus amyloliquefaciens subtilisin DFE precursor有99%的同源性,对应的氨基酸序列与Bacillussp.DJ-4有99%的同源性。apr基因在大肠杆菌BL21中获得表达,并表现出蛋白酶活性。结论:获得了具有活性的新的碱性蛋白酶基因。

Objective：To acquire gene encoding alkaline protease.Methods： Gene fragment encoding alkaline protease was amplified by PCR from Bacillus subtilis A-109,sequenced and analyzed.The recombinant plasmid was constructed and transformed into E.coli BL21.SDS-PAGE detected the expression of the gene fragment.Results：The gene fragment is 1092 bp.The nucleotides sequence homology and putative amino acid sequence homology of the gene fragment were 99%,compared with Bacillus amyloliquefaciens subtilisin DFE precursor and Bacillus sp.DJ-4 ,respectively. The apr gene was expressed in E. coli BL21 and showed its activities of protease. Conclusion： A new gene enceding alkaline protease was obtained.