犬瘟热病毒疫苗株与野毒株H蛋白球状头部域的免疫原性比较研究  

A Comparison of the Preliminary Immunogenicity Between the Head Domain in H Protein of vaccine and that of wild strain of Canine Distemper Virus

作　　者：秦晓冰[1] 王怡男[1] 张传美[1] 黄娟[1] 张洪亮[1] 王聪[1] 胡琳琳[2] 单虎[1] 

机构地区：[1]青岛农业大学动物科技学院山东省预防兽医学重点实验室,山东青岛266109 [2]山东省安丘市畜牧局,山东安丘262100

出　　处：《中国兽医杂志》2015年第6期26-29,共4页Chinese Journal of Veterinary Medicine

基　　金：国家863计划(2011AA100600);公益性行业(农业)科研专项(201303042);科技部畜禽重要疫病流行病学调查科技基础性工作专项(2012FY111000)

摘　　要：以犬瘟热病毒疫苗株CDV3和野毒株LYM基因组RNA为模板,RT-PCR扩增得到CDV3和LYM的H全基因序列,PCR分别扩增得到CDV3和LYM-Head Domain基因,将其分别克隆于p ET-30a载体进行诱导表达,经SDS-PAGE和Western Blot分析表明,这两种重组蛋白均正确表达,分子质量约为50.8 ku。再将两种重组蛋白分别免疫小鼠,间接ELISA结果表明,这两种重组蛋白均能激发机体产生特异性抗体,具有一定免疫原性。中和试验结果表明,这两种重组蛋白的抗体中和效价差异不显著(P>0.05)。All of genome RNAs were extracted from CDV3/LYM strains of canine distemper（CD ）, and H genes were obtained by RT-PCR. Accordingly, Head Domain genes were amplified by PCR from H gene and subcloned into pET-30a-c （＋）to construct recombinant plasmids, named as pET-30a-Head Domain, then the two recombinant plasmids were transformed to the competence E.coli BL21 （DE3）for inducible expression, The target proteins could be identified by SDS-PAGE and Western Blot analyses. Sera from mice inoculated by the above proteins were monitored through indirect ELISA. It has manifested that the two proteins could equally produce specific antibodies and have similar immunogenicity. There were no significant differences between antibody titers of the two target proteins according to neutralization test （P〉0.05）. This study contributed in locating neutralizing epitopes of H proteins and discerning their humoral immune mechanisms.

关 键 词：犬瘟热病毒 H蛋白 球状头部域 原核表达 免疫原性 

分 类 号：S852.651[农业科学—基础兽医学]