【摘要】 枯草芽孢杆菌转换效率低一直制约着该菌基因改造技术的应用,本研究比较了化学法和高渗电转化法对枯草芽孢杆菌转化率的影响,发现高渗电转化法的转化效率较高。从Bacillus alcalophillus PB92中扩增出碱性蛋白酶基因Mapr,构建成重组分泌型表达载体pWB980-Mapr,碱性蛋白酶基因在枯草芽孢杆菌DB104中得到表达。SDS-PAGE分析,重组蛋白酶的分子量为28000。pWB980-Mapr在枯草芽孢杆菌中表达的酶活为1563 U/ml。更多还原

【Abstract】 The application of genetical modification technology has been constrained by low transformation efficiency in Bacillus subtilis.Compared with chemical transformation,the hypertonic electroporation exhibited higher transformation efficiency in this experiment.The sequence of alkaline protease gene Mapr cloned from Bacillus alkalophillus PB92 chromosomeal DNA was inserted into B.subtilis vector pWB980 to yield the recombinant plasmid pWB980-Mapr.After the transformaiton by hypertonic electroporation,pWB980-Mapr was expressed in the host B.subtilis DB104.SDS-PAGE analysis showed a molecular weight of 28 000 and an activity of 1 563 U/ml of recombinant protease.更多还原