乌苏里貉IgG Fc片段克隆表达及免疫交叉源性分析  

Expression and analysis of immunity cross-reaction on racoon dog IgG-Fc

作　　者：赵翠[1] 刘玲红[1] 郭树源[1] 庞晓茹 宋道祯 付石军 常维山[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]山东省滨州兽医研究所,山东滨州256600

出　　处：《中国兽医学报》2016年第11期1864-1868,1890,共6页Chinese Journal of Veterinary Science

基　　金：山东省毛皮动物创新团队滨州综合试验站项目(SAT-180-011-15)

摘　　要：为研究乌苏里貉IgG Fc与犬IgG间的免疫交叉源性,本研究采用斑点酶联免疫吸附法(dot-ELISA)初步确定犬与乌苏里貉血清抗体间存在较强免疫交叉反应源性;通过RT-PCR成功克隆乌苏里貉IgG Fc片段基因,构建pcDNA3.1-IgG(Fc)真核表达质粒,转染HEK293T细胞,借助间接免疫荧光试验(IFA))证实该蛋白成功表达(大小约为22 000),免疫印迹(Western blot)试验证明该蛋白与抗犬IgG存在免疫结合反应。结果表明,乌苏里貉IgG Fc与犬IgG间存在强烈的免疫交叉源性,应用犬源高免血清或犬免疫球蛋白治疗貉犬瘟热具有可行性,同时为今后构建特定抗原基因与IgG Fc基因融合表达载体,研究DNA疫苗及Fc的生物佐剂提供了理论依据。To study the immunity cross reaction between racoon dog IgG-Fc and dog IgG, dot- ELISA, RT-PCR, eukaryotic expression, IFA and Western blot were carried on. It was proved that there was strong immunity cross-reaction between serum antibody of racoon dog and anti-dog antibody by means of dot-ELISA. The eukaryotic expression vector pcDNA3.1-IgG（Fc） ,which was successfully constructed,was expressed in HEK 293T cell. And the protein had the strong cross reaction to the HPR-rabbit aganist dog antibody. All these will not only build an important theoretical foundation on whether antiserum for dog can be used for treating the CDV of raccoon dog, but also provide an experimental basis on DNA vaccine and biological adjuvant about Fc gene.

关 键 词：乌苏里貉 免疫球蛋白(G)Fc 真核表达 免疫交叉反应 犬瘟热 

分 类 号：S865.24[农业科学—野生动物驯养]