犬瘟热病毒基因疫苗表达载体构建及在真核细胞中的表达

Construction of expressing gene vaccine vectors of canine distemper virus and expression in MDCK cells

利用pCI载体构建了犬瘟热病毒基因疫苗表达载体质粒pCIF和pCIN,通过脂质体介导法将这两种真核表达质粒分别转染MDCK细胞,用RT-PCR法进行转录水平的检测,并用间接ELISA法检测目的蛋白是否表达。结果表明,真核表达质粒pCIF和pCIN已被成功构建,这两种重组质粒转染MDCK细胞72h后就可检测到目的基因的转录和两种目的蛋白的表达,表达的蛋白均能与抗CDV抗体发生特异性的抗原抗体反应。这为研究预防犬瘟热的基因疫苗奠定了良好的基础。

Two expressing vectors plasmids pClN and pCIF were successfully constructed with pCI vector. After pCIF and pCIN plasmid vectors were transfected into MDCK cells with Lipofectamine, transcriptions of targeted genes were detected by RT-PCR and the expressing objective proteins were detected by ELISA method. The results showed that the transcription of objective genes and expression of interesting proteins could be detected at 72 h post-transfection, respectively, The special antigen-antibody reaction was detected between the expressing protein and antibody of CDV. This was a base on studying gene vaccine of CDV.