中国畜牧兽医2016，Vol.43Issue(8)：1938-1943,6.DOI:10.16431/j.cnki.1671-7236.2016.08.02
鸡 MHCⅠα和β2 m链基因真核表达载体的构建及其在293T 细胞中的表达
Construction of Eukaryotic Expression Vector of Chicken MHCⅠα andβ2 m Genes and Its Expression in 293T Cells
摘要
Abstract
To explore the mechanism of MHC Ⅰ molecule in immune response ,chicken MHCⅠαandβ2 m genes were cloned by PCR .Then the fragments were inserted into the eukaryotic expres‐sion vector with fluorescent protein ,and the recombinant plasmids pEGFP‐M HCⅠαand pmCher‐ry‐M HCⅠβ2 m were constructed .The recombinant plasmids were transfected into 293T cell with lipofectin reagent .The gene products of recombinant plasmids were mainly located to endomem‐brane system of the cells by fluorescence microscopy ,and changed the intracellular localization of the fusion with the fluorescent protein .Moreover ,the positive reactions were observed by the method of Western blotting ,and the proteins had the molecular weight of 68 .3 and 41 .3 ku ,re‐spectively ,in accord with the target proteins .The results showed that the recombinant plasmids were expressed in 293T cells with a good immunological activity ,and the proteins had the binding reaction with specific antibodies .关键词
鸡MHCⅠα基因/鸡MHCⅠβ2 m链基因/真核表达载体/293T细胞Key words
chicken M HC Ⅰ α gene/chicken M HC Ⅰ β2 m gene/eukaryotic expression vector/293 T cell分类
生物科学引用本文复制引用
戴银,胡晓苗,赵瑞宏,侯宏艳,沈学怀,潘孝成,周学利,张丹俊..鸡 MHCⅠα和β2 m链基因真核表达载体的构建及其在293T 细胞中的表达[J].中国畜牧兽医,2016,43(8):1938-1943,6.基金项目
国家自然科学基金项目（31302044）；国家蛋鸡产业技术体系项目（CARS-41）；安徽省科技攻关项目 （）
