人纤溶酶原K1-3区基因在大肠杆菌中的表达  

Efficient Expression of Human Plasminogen Kringle 1-3 Domain Gene in E.coli

作　　者：陆幸妍[1] 盛节[1] 张添元[1] 罗进贤[1] 

机构地区：[1]中山大学基因工程教育部重点实验室,广东广州510275

出　　处：《中山大学学报（自然科学版）》2001年第3期63-65,共3页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家自然科学基金!资助项目 (396 70 0 13) ;广东省自然科学基金!资助项目 (96 0 0 5 2 )

摘　　要：将人纤溶酶原kringle 1- 3 (K1- 3)基因插入融合表达载体pET - 17b ,获得重组质粒pET -K13 ,转化E coliBL2 1(DE3) ,在IPTG诱导下 ,人纤溶酶原K1- 3基因在E coliBL2 1(DE3,pET -K13)中获得高效融合表达 ,表达量占菌体总蛋白的 2 4% ,表达产物以包涵体存在 ,Westernblot证明重组蛋白对人纤溶酶原抗血清有特异免疫原性 .The kringle 1-3 domain(K1-3) of plasminogen is a potent inhibitor of endothelial cell proliferation and angiogenesis.The gene expression of the K1-3 domain of human plasminogen in \%E\^coli\% is reported. The K1-3 gene was first inserted into the expression vetor pET-17b,resulting in plasmid pET-K13.pET-K13 was then transformed into \%E\^coli\% BL21(DE3).The transformant \%E\^coli\% BL21(DE3,pET-K13) was used to detect the gene expression.SDS_PAGE result showed that the K1-3 gene was efficiently expressed in \%E\^coli\% BL21(DE3,pET-K13) by IPTG induction.The expressed product constitutes 24% of the total bacterial protein as estimated by densitometer and exists as inclusion body.Western blot result indicates that the recombinant protein reacts with antiserum against human plasminogen.

关 键 词：人纤溶酶原K1-3 基因表达 大肠杆菌 WESTERN BLOT 肿瘤 血管生成抑制素 

分 类 号：Q786[生物学—分子生物学] R730.2[医药卫生—肿瘤]