鸡MHCⅠ分子单链三聚体真核表达载体的构建和表达  

Construction and Expression of Single Chain Trimer of Chicken MHCⅠMolecular

作　　者：戴银[1] 单文杰[2] 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] DAI Yin SHAN Wen-jie HU Xiao-miao ZHANG Dan-jun et al(Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Science, Hefei, Anhui 230031 College of Animal Science and Technology, Anhui Agricultural University, Hefei, Anhui 230036)

机构地区：[1]安徽省农业科学院畜牧兽医研究所,安徽合肥230031 [2]安徽农业大学动物科技学院,安徽合肥230036

出　　处：《安徽农业科学》2017年第1期139-142,共4页Journal of Anhui Agricultural Sciences

基　　金：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(11010302119)

摘　　要：[目的]探索主要组织相容性复合体Ⅰ类分子(Major Histocompatibility Complex,MHCⅠ)提高抗原呈递效率的方法,获得具有免疫学活性的抗原肽-MHCⅠ分子单链三聚体。[方法]采用基因重组技术,通过连接肽(G4S)3将鸡新城疫病毒抗原表位短肽HN(aa346-353)与鸡MHCⅠ(B-F)分子二聚体的编码基因共价串联,构建带有绿色荧光标签蛋白的真核表达载体p EGFP-HN_(346-353)-B-Fβ2m-α。[结果]转染真核细胞293T后,荧光显微镜观察可见,重组融合蛋白HN_(346-353)-B-Fβ2m-α分子三聚体主要表达定位于真核细胞的内膜系统。Western Blot结果表明,在预期蛋白分子量大小位置有明显的特异反应带,重组蛋白与其相应抗体可发生特异性结合反应。[结论]重组质粒能够在真核细胞中表达,且融合蛋白具有良好的生物学活性。[Objective] To improve the efficiency of antigen presentation of MHC I molecule, the single chain trimer of pepide-MHC I mole- cules were obtained. [ Method] The chicken MHC I （B-F） gene and the epitope protein HN （aa346-353） of Newcastle disease virus were linked by a linker with sequences encoding（ G4S） 3. Then the recombinant plasmids （pEGFP-HN346-353-B-F β2m-α） with green fluorescent protein was constructed. Further the recombinant plasmid was transfected into eukaryotic cell 293T. [ Result] The gene product of recombinant plasmid was mainly located to endomembrane system of the cells by fluorescence microscopy. Moreover, the positive reactions were observed by the method of Western Blot, in accordance with the target proteins, and the proteins had the binding reaction with specific antibodies. [Conclusion] The recombinant plasmid was expressed in eukaryotic cells with a good biological activity.

关 键 词：鸡MHC B-F分子 单链三聚体 真核表达 

分 类 号：S188.2[农业科学—农业基础科学]