地衣芽孢杆菌耐高温α-淀粉酶基因在大肠杆菌中的克隆及表达

Cloning and Expression of the Gene Encoding Heat-stable α-amylase from Bacillus licheniformis in E.coli JM109

用PCR方法从地衣芽孢杆菌中扩增了耐高温α淀粉酶基因,将扩增的DNA片段插入到大肠杆菌载体pUC19中,构建重组分泌型表达载体pUAM。pUAM中的耐高温α淀粉酶基因在大肠杆菌JM109中得到表达。经SDSPAGE分析显示,蛋白表达产物的分子量为55ku,同核酸序列测定所推导的值相符。

The heat-stable α-amylase gene was amplified from Bacillus licheniformis by using PCR technique.The amplified 1.9kb DNA fragment was inserted into an E.coli vector pUC19 to yield the recombinant secrete plasmid pUAM.The heat-stable α-amylase gene in Bacillus licheniformis was expressed in E.coli JM109.SDS-PAGE analysis shows that the molecular weight of the expressed recombinent product is about 55 ku,which correspond with the prediction by gene sequence.