口蹄疫病毒二价DNA疫苗的构建及实验免疫研究

通过PCR方法从本室已构建的克隆载体pGEMT-P1-2A获得O型口蹄疫病毒主要保护性抗原VP1基因，以基因突变获得A型VP1基因。将这两种血清型的VP1基因串联后连接到真核表达载体PVAX1 PCMV启动子下游，构建成口蹄疫二价核酸疫苗pVAX1-OA。经Western blot和IFA检测，目的蛋白在HeLa细胞中获得正确表达。动物实验表明，免疫小鼠T淋巴细胞明显增殖，特异性CTL杀伤活性较对照组显著提高；血清抗体能分别与O型和A型抗原反应，抗体效价均高于空白对照组，但较灭活苗低。

Construction of Bivalent DNA Vaccine Against Foot-and-Mouth Disease and Detection of Immunitical Activity

The major epitope VP1 gene of serotype O of foot and mouth disease virus (FMDV) was obtained from clone vector pGEMT P1 2A by PCR. Serotype A VP1 gene was obtained by mutation. The genes of the two serum types were connected in series and then linked to the downstream of the promoter of eukaryotic expression vector pVAX1 to construct bivalent DNA vaccine pVAX1 OA against FMDV. The protein of interest was expressed in HeLa cell line as confirmed by Western blot and IFA. The experiment in vitro showed that T lymphocytes of the immunized mice proliferated significantly (P<0 01) and the specific cytotoxic T lymphocyte (CTL) activity was obviously higher than that of the controls. The antiserum could react with serotype O and serotype A antigens. The titer of the antiserum was both higher than that of the controls, but it was still much lower than that of the inactivated vaccine.