赵宇军1，朱远茂2，相文华2
（1.山西农业大学 动物科技学院, 山西 太谷030801; 2.中国农业科学院 哈尔滨兽医研究所, 黑龙江 哈尔滨050001）
摘 要：设计引物，含有kozak序列及BamHI的的上游引物和带有终止密码子及EcoRV酶切位点的下游引物，以猪HEV DQ1 ORF2为模板，进行PCR。将扩增片段和pcDNA3.1质粒以BamHI/EcoRV进行双酶切后进行连接。连接产物转化至大肠杆菌DH5α，经测序证明该序列正确，命名为pcDQ1。进行pcDQ1质粒提取，以vero细胞为表达细胞进行转染，以间接免疫荧光试验进行验证。以100 μg /次/只剂量的pcDQ1对BAL B/C小鼠进行免疫以获取单因子血清。共免疫3次，采集血清，进行ELISA效价测定。结果表明，该核酸疫苗可以免疫使小鼠产生抗体。
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Construction of DNA Vaccine for Swine Hepatitis E Virusand Immunology Effect on Mice
ZHAO Yu-jun1，ZHU Yuan-mao2，XIANG Wen-hua2
(1．College of Animal Science of Shanxi Agriculture University , Taigu 030801, Shanxi, China; 2.Haerbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Haerbin 050001, Heilongjiang, China)
Abstract: Using swine HEV DQ1strain ORF2 as template, and a pair of primer with kozak sequence , by PCR we got whole ORF2 of swine HEV DQ1. By Inserting the amplified part into vaccine vector pcDNA3.1, we constructed the DNA vaccine for swine HEV DQ1. After translated DNA vaccine into vero cell and immunized BAL B/C mice ,we found that the vaccine could produce protein detected by immunofluorscence assay in vero cells and induce immunology reaction in mice.
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