摘要
将含有kozak序列及BamH I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HEVDQ1 ORF2为模板,进行PCR。将扩增片段和pcDNA3.1质粒以BamH I/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100μg/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。
Using swine HEV DQ1 strain 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.
出处
《激光生物学报》
CAS
CSCD
2006年第3期301-304,共4页
Acta Laser Biology Sinica
基金
中国农科院院长基金(2003168)
