谷氨酰胺转胺酶基因的融合表达  

Expression of Transglutaminase Fusion Protein

作　　者：黄柯[1,2] 金志华[1,2] 金庆超[1] 洪小伟[2] 郑喜[2] 

机构地区：[1]浙江大学宁波理工学院生物与化学工程分院,浙江宁波315100 [2]浙江大学化学工程与生物工程系,浙江杭州310027

出　　处：《广州化工》2010年第6期97-100,共4页GuangZhou Chemical Industry

摘　　要：运用基因工程手段在大肠杆菌中高效表达谷氨酰胺转胺酶基因。以茂原链霉菌基因组DNA为模板,通过PCR扩增得到一条长为1224bp的谷氨酰胺转胺酶基因片段,成功构建原核表达载体pGEX-TGase,并转化大肠杆菌BL21(DE3)获得重组菌BL21/pGEX-TGase。重组菌经IPTG诱导产生GST-TGase融合蛋白,并对表达条件进行了优化,应用亲和层析方法纯化融合蛋白,对纯化产物进行SDS-PAGE电泳分析,结果显示目的蛋白获得了较高水平的表达。A recombinant bacterium was obtained,which had high level expression of transglutaminase.A 1224bp of transglutaminase gene fragment from streptomyces mobaraensis was obtained by PCR,and then was inserted into multi-cloning site of pGEX-6p-1 vector.The recombinant plasmid was then transformed into E.coli BL221（DE3） producing the recombinant BL21/pGEX-TGase.GST-TGase fusion protein was induced by IPTG,and the induction conditions were optimized.The fusion protein was absorbed on the Glutathione Sepharose.After cutting the fusion protein by thrombin,TGase can be purified.The result of SDS-PAGE revealed the recombinant protein had a high level expression.

关 键 词：谷氨酰胺转胺酶 pGEX 融合蛋白 

分 类 号：Q78[生物学—分子生物学]