鸡MHCⅠ分子二聚体融合基因的构建及表达  

Construction and Expression of Fusion Gene of Chicken MHCⅠ Dimers

作　　者：戴银[1] 胡晓苗[1] 赵瑞宏[1] 侯宏艳[1] 沈学怀[1] 潘孝成[1] 周学利[1] 张丹俊[1] 

机构地区：[1]安徽省农业科学院畜牧兽医研究所,合肥230031

出　　处：《中国农学通报》2016年第20期1-5,共5页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金项目"MHCⅠ分子靶向性疫苗载体的构建及其免疫作用的机理"(31302044);国家现代农业产业技术体系专项"合肥综合试验站"(CARS-41);安徽省科技攻关项目"鸡J-亚群白血病综合防治技术研究"(11010302119)

摘　　要：为进一步研究主要组织相容性复合体Ⅰ类分子(major histocompatibility complex,MHCⅠ)的作用机制,提高其抗原呈递的效率,获得生物活性较好的鸡MHCⅠ分子二聚体,采用4个不同长度的连接肽G_4S、(G_4S)_2、(G_4S)_3和(G_4S)_4,通过基因重组技术将MHCⅠα和β2m链基因共价连接,随后插入带有绿色荧光标签蛋白的真核表达载体,成功构建了4个重组质粒pEGFP-MHCⅠβ_2m-α。真核细胞表达后,荧光显微镜观察可见,各重组MHCⅠ二聚体分子均主要分布于真核细胞的内膜系统。此外,Western Blot检测均可见蛋白大小约80.0 k D左右的特异反应带,表达蛋白与鸡MHCⅠ类分子的相应抗体可发生特异性结合反应。研究结果表明各连接肽的重组质粒p EGFP-MHCⅠβ_2m-α均能够在真核细胞中顺利表达,且融合蛋白具有良好的免疫学活性。To explore the mechanism of major histocompatibility complex class Ⅰ（MHC Ⅰ）, improve theefficiency of antigen presentation, the dimers of chicken MHC molecules were obtained. The MHC Ⅰ α andβ_2m chain gene were linked by different length linkers with sequences encoding G_4S,（G_4S）_2,（G_4S）_3 and（G_4S）_4.Then this fragments were inserted into the eukaryotic expression vector with green fluorescent protein, and therecombinant plasmids（p EGFP-MHCⅠβ_2m-α） were constructed. The recombinant plasmids were transfectedinto eukaryotic cells. The gene products of recombinant plasmids were mainly located at endomembrane systemof the eukaryotic cells under fluorescence microscopy. Moreover, the positive reactions were observed byWestern Blot, and all the proteins had a molecular weight of about 80.0 k D, in accordance with the targetproteins, and the proteins had binding reaction with specific antibodies. The results showed that therecombinant plasmids with different length linkers could be expressed in eukaryotic cells with a goodimmunological activity.

关 键 词：鸡MHCⅠ分子 二聚体 真核表达 

分 类 号：S852[农业科学—基础兽医学]