人组织型谷氨酰胺转氨酶基因的克隆 表达及纯化  

Gene Cloning,Expression,and Purification of Human Transglutaminase 2

作　　者：麻丽霞[1] 曹文君[1] 

机构地区：[1]长治医学院心血管病研究所,046000

出　　处：《长治医学院学报》2015年第6期405-408,共4页Journal of Changzhi Medical College

基　　金：长治医学院科技创新团队资助项目(CX201401)

摘　　要：目的:克隆人组织型谷氨酰胺转氨酶基因,并进行原核表达及纯化。方法:从Caco-2细胞中提取总RNA,利用RT-PCR克隆人组织型谷氨酰胺转氨酶基因,然后将其连接到原核表达载体pET-32a。将构建成功的重组表达载体pET-32a-TG2导入大肠杆菌BL21(DE3),IPTG诱导表达His-TG2融合蛋白。用Ni-NTA蛋白纯化柱对融合蛋白进行纯化,采用Western Blotting检测重组蛋白的特异性。结果:成功克隆出人组织型谷氨酰胺转氨酶基因,构建了pET-32a-TG2原核表达载体,并成功诱导了重组融合蛋白His-TG2的表达。结论:构建了表达人组织型谷氨酰胺转氨酶基因的原核表达系统,并成功对重组蛋白进行了纯化。Objective：To study the gene cloning,expression,and purification of human Transglutaminase 2.Methods：Total RNA was extracted from Caco-2cells,and human transglutaminase 2 gene was amplified by RT-PCR.The target fragment was inserted into vector pET-32 a,and the recombinant vector pET-32a-TG2 was transformed and expressed in E.coli strain BL21（DE3）.The His-TG2 fusion protein was purified by Ni-NTA affinity chromatography.Results：The human transglutaminase 2 gene was cloned and inserted into expression vector pET-32 a.The fusion protein His-TG2 was expressed and purified with high specificity.Conclusion：This study constructed a prokaryotic expression system of human transglutaminase2 gene,and the recombinant His-TG2 fusion protein was successfully purified.

关 键 词：人组织型谷氨酰胺转氨酶基因 原核表达 蛋白纯化 

分 类 号：R392[医药卫生—免疫学]