纳豆激酶基因的克隆及表达研究  

Cloning and Expression of the Nattokinase Gene

作　　者：姜媛媛 王曙文 王铁东[2] 丁东红[3] 王景会[1,4] 

机构地区：[1]中国农业科技东北创新中心农产品加工研究中心,长春130033 [2]吉林大学农学部,长春130062 [3]吉林大学第四医院-一汽总医院,长春130011 [4]吉林农业大学食品科学与工程学院,长春130118

出　　处：《吉林农业大学学报》2009年第4期398-401,共4页Journal of Jilin Agricultural University

基　　金：吉林省科技发展计划项目(20070579)

摘　　要：经PCR扩增获得了纳豆激酶基因(NK)。将此基因插入原核表达载体pTWIN1中,使之与几丁质结合域(CBD)一内含肽(intein)融合,获得原核表达质粒pTWIN1/NK。转化宿主菌E.coliBL21(DE3),在IPTG诱导下进行纳豆激酶的表达研究。SDS-PAGE电泳分析结果表明,重组蛋白在BL21(DE3)中获得高效表达,在低温诱导时主要以可溶性蛋白的形式存在。Nattokinase gene was amplified using polymerase chain reaction （PCR）, and cloned into prokaryotic expression vector pTWIN1. The expression plasmids of pTWIN1/NK was constructed and was further transformed into E. coli BL21 （DE3）. Expression of the enzyme was performed at different temperatures with IPTG induction. SDS - PAGE showed that nattokinase was expressed in soluble form at a low temperature. The cloning and expression of nattokinase would be helpful in producing nattokinase by gene engineering strain.

关 键 词：纳豆激酶基因 克隆 表达 

分 类 号：Q786[生物学—分子生物学]