纳豆激酶基因的克隆与表达  

Cloning and Expression of Nattokinase Gene

作　　者：张淑梅[1] 张云湖[1] 赵晓祥[1] 李晶[1] 金红星[1] 田洁萍[1] 

机构地区：[1]黑龙江省科学院应用微生物研究所,哈尔滨150010

出　　处：《中国生物化学与分子生物学报》1999年第6期912-915,共4页Chinese Journal of Biochemistry and Molecular Biology

基　　金：黑龙江省自然科学基金

摘　　要：利用ＰＣＲ方法从分泌纳豆激酶的枯草杆菌基因组ＤＮＡ 中扩增得到了纳豆激酶基因，并测定其核苷酸序列．利用基因重组技术构建了纳豆激酶基因的表达载体，并在大肠杆菌中进行了表达．ＳＤＳ－聚丙烯酰胺凝胶电泳表明，表达蛋白占菌体蛋白的１５．２％ ，琼脂糖－纤维蛋白平板法测出表达产物具有溶解血栓活性．Nattokinase is a kind of fibrinolytic enzyme.Compared with other fibrinolytic enzymes,it has many merits,such as the longer time in vivo, no harm to people.DNA fragment encoding nattokinase was amplified by PCR from genomic DNA,and cloned into pUC19 vector,then sequenced.The expression vector of fusing nattokinase gene was constructed by gene recombination technology.As a result,high level expressior E.coli strain of nattokinase was obtained.It was proved that expression protein was 15.2% of total cell protein by SDS PAGE and it had the activity of dissolving thrombus.

关 键 词：纳豆激酶基因 序列 表达 克隆 溶栓药 

分 类 号：TQ464.8[化学工程—制药化工] R972[医药卫生—药品]