利用血红蛋白基因提高短小芽孢杆菌在低氧条件下的碱性蛋白酶产量  

Improving production of alkaline protease using hemoglobin gene expression in Bacillus pumilus

作　　者：张风豪[1] 王海燕[1] 何明雄[1] 李羚锐[1] 杨春晖[1] 葛建[1] 张义正[1] 

机构地区：[1]四川大学生命科学学院,四川省分子生物学及生物技术重点实验室,成都610064

出　　处：《高技术通讯》2007年第7期755-760,共6页Chinese High Technology Letters

基　　金：863计划（2004AA214111）和四川省＂十五＂重点攻关（02NG002-005）资助项目.

摘　　要：将透明颤菌血红蛋白基因vgb成熟肽编码区序列分别与枯草芽孢杆菌（Bacillus subtilis）P43双启动子和短小芽孢杆菌（B. pumilus）碱性蛋白酶基因启动子wBp连接,利用大肠杆菌-枯草芽孢杆菌穿梭载体pSUGV4分别构建组成型表达质粒pSUP43Vgb和pSUwBpVgb,将其分别转化短小芽孢杆菌UN-31-C-11.SDS-PAGE和CO差异光谱检测表明,融合基因在UN-31-C-11中均得到了表达,并且表达质粒使重组菌株在低氧条件下的生长量在对数生长后期超过对照菌株20%以上,同时促进了短小芽孢杆菌碱性蛋白酶基因的表达,其产量高于对照30%以上.Two recombinant plasmids were constructed by inserting the mature peptide-coding sequence of the Vitreoscilla hemoglobin gene （vgb） to the downstream of the Bacillus subtilis promoter P43 and B. pumilus alkaline protease gene promoter wBp respectively, and then to the shuttle vector. The recombinant plasmids were called pSUP43Vgb and pSUwBpVgb. These expression plasmids were transformed into B. pumilus UN-31-C-11. SDS-PAGE and carbon monoxide binding assay demonstrated that Vitreosciua hemoglobin （VHb） gene was expressed in UN-31-C-11. In B. p umilus, cells containing recombinant expression plasmids had the cell density 20 % higher than the control in the late exponential phase. Meanwhile, recombinant plasmids could make the alkaline protease production 30% higher than the control.

关 键 词：透明颤菌血红蛋白基因(vgb) 基因启动子 穿梭载体 短小芽孢杆菌 基因表达 

分 类 号：Q78[生物学—分子生物学]