带自身启动子的bgaB基因在枯草芽孢杆菌中的表达  

The Expression of bgaB Gene with its Promoter in Bacillus subtilis

作　　者：傅晓燕[1] 陈卫[1] 夏雨[1] 张灏[1] 

机构地区：[1]江南大学食品科学与安全教育部重点实验室,江苏无锡214036

出　　处：《食品与生物技术学报》2005年第3期44-47,51,共5页Journal of Food Science and Biotechnology

摘　　要：将来源于嗜热脂肪芽孢杆菌的启动子连同β-半乳糖苷酶bgaB基因经PCR扩增后,连接在T载体上,再取代枯草杆菌载体pZ01-bgaB的启动子,将其在枯草杆菌宿主WB600中表达.经摇瓶发酵20h,得到乳糖酶活力6.37U/mL,比活力3.814U/mg,SDS-PAGE电泳显示有明显重组蛋白质条带.证明了嗜热脂肪芽孢杆菌来源的启动子在枯草杆菌中是完全适用的.The promoter and gene sequence from Bacillus stearothermophilus were amplified through PCR, and cloned into T-vector. Then bgaB gene and its regulatory sequence were constructed into B. subtilis vector pZ01-bgaB to replace the original promcter. After culturing for 20 h, thermostable β-galactosidase activity of 6. 37 U/mL, and specific activity of 3. 814 U/ mg were obtained. The target protein was identified at about 70 kD in SDS-PAGE. The result suggested that the bgaB promoter from B. stearothermophilus was effective in B. subtilis.

关 键 词：嗜热脂肪芽孢杆菌 启动子 枯草芽孢杆菌 6gαB基因 

分 类 号：Q786[生物学—分子生物学]