犬组织相容复合物Ⅰ类分子的真核表达和细胞膜呈递  

Over-expression and surface present of canine major histocompatibility complex class Ⅰ in FK81 cell line

作　　者：贺英[1] 陈娟[1] 潘素敏 倪静[1] 曹旺斌[1] 杨宗泽[1] 史秋梅[1] HE Ying;CHEN Juan;PAN Su-min;NI Jing;CAO Wang-bin;YANG Zong-ze;SHI Qiu-mei(Hebei Key Laboratory of Preventive Veterinary ,Department of Animal Science, Hebei Normal University of Science ＆ Technology ,Qinhuangdao,Hebei 066600,Chin)

机构地区：[1]河北科技师范学院动物科技学院河北省预防兽医学重点实验室

出　　处：《中国兽医学报》2018年第6期1132-1135,共4页Chinese Journal of Veterinary Science

基　　金：河北省自然科学基金资助项目(C2013407106);国家星火计划资助项目(2015GA620002);特种毛皮动物高效健康养殖关键技术集成应用与示范资助项目

摘　　要：为进一步研究主要组织相容性复合体Ⅰ类分子（major histocompatibility complexclass Ⅰ，MHCⅠ）的作用机制，提高其抗原呈递效率，通过基因重组技术获得犬DLA-12基因，插入pcDNA3真核表达载体，成功构建了重组质粒pcDNA3-DLA-12。用重组质粒脂质体法转染FK81细胞，G418筛选稳定转染细胞，获得表达MHCI克隆细胞。Western blot检测可见43000左右的蛋白特异反应带，与目的蛋白大小相当。流式细胞仪分析表明，超表达DLA-12增加了MHCⅠ的细胞膜呈递。To explore the mechanism of major histocompatibility complex class Ⅰ （MHC Ⅰ ） and improve the efficiency of antigen presentation,the cainine MHC Ⅰ DLA-12 were obtained. Then the gene fragment was inserted into the eukaryotic expression vector, and the recombinant plasmid （pcDNA3-DLA-12） was constructed. The recombinant plasmid was transfected into FK81 line. The over-expression DLA-12 cell was obtained by G418 seletion. The positive reactions were observed by Western blot, and the proteins had a molecular weight of about 43 000, in accordance with the target proteins. Surface expression of MHC I was investigated in over-expression DLA- 12 cells by flowcytometry using the MHC I specific monoclonal antibody（mAb） w6/32 and the protein expression on cell surface was increaed. The cell were inoculated with canine parvovirus, comparison with control FK81 cell, the repliation of virus was decreased significantly in over-expression DLA-12 cell. Overexpression MHC Ⅰ increased its present on cell surface and inhibited virus replication.

关 键 词：犬 主要组织相容性复合体Ⅰ类分子 真核表达 细胞膜呈递 

分 类 号：S852.42[农业科学—基础兽医学]