摘要
采用PCR技术 ,以基因组DNA为模板克隆得到运动发酵单胞菌 (Zymomonasmobilis)乙醇脱氢酶 (alcoholdehydrogenaseⅡ )基因adhB ,连接到表达载体 pSE380上 ,得到重组质粒 pSE adhB。将此重组质粒转化到大肠杆菌菌株DH5α中 ,重组菌株经IPTG诱导后 ,在乙醛指示平板检测到乙醇脱氢酶活性。SDS PAGE电泳结果显示出明显的 4 0KD特异性蛋白质条带。重组菌株经诱导培养 ,每毫升发酵液酶活力为 5u。
The gene adhB encoding alcohol dehydrogenaseⅡ(ADHⅡ)of Zymomonas mobilis was amplified by PCR from total DNA . The recombinant plasmid,pSE-adhB was constructed by inserting adhB into expression vector pSE380 and then transformed E.coli DH5α. The recombinant strains were induced by IPTG to express ADHⅡ. The activity was determined by aldehyde indicator plates. Expression level of recombinant alcohol dehydrogenaseⅡ(ADHⅡ) reached 5u/mL.By companision, no activity appeared in hosted E.coli DH5α.
出处
《工业微生物》
CAS
CSCD
北大核心
2004年第1期17-20,共4页
Industrial Microbiology
基金
国家"8 63"资助项目 (编号 2 0 0 1AA2 14 171)
