口蹄疫病毒P1-2A和3C蛋白重组腺病毒的构建及其免疫原性研究  

Recombinant adenovirus expression of FMDV P1-2A and 3C protein and its immune response in mice

作　　者：张春红[1] 郭春和[1] 项林盛[1] 唐丽云[1] 郑红玉[1] 姜后全[1] 黄毓茂[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国预防兽医学报》2012年第6期480-484,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：广东省现代农业生猪产业技术体系(F10021)

摘　　要：为研制口蹄疫病毒(FMDV)的新型重组腺病毒疫苗,本研究通过RT-PCR扩增FMDV GD株3C、P1、P1-2A、L-P1和L-2A基因,分别构建重组腺病毒穿梭质粒,并在含有腺病毒骨架质粒pAdEasy-1的BJ5183E.coli中同源重组获得腺病毒重组质粒pAd-3C、pAd-P1、pAd-P1-2A、pAd-L-P1、pAd-L-2A,经PacⅠ线性化后转染AD-293细胞,获得含有目的基因的重组腺病毒。将具有感染能力的复制缺陷型重组腺病毒rAd-3C分别与rAd-P1、rAd-P1-2A、rAd-L-P1、rAd-L-2A共感染Vero细胞,裂解细胞收集细胞上清液并进行小鼠免疫试验。经ELISA检测表明,重组腺病毒rAd-3C分别与rAd-P1-2A、rAd-L-2A共感染Vero细胞上清液能够诱导小鼠产生特异性体液免疫应答。To develop a recombinant virus vaccine against foot-and-mouth disease （FMD）, the 3C, P1, P1-2A, L-P1 and L-2A genes of FMDV were amplified by RT-PCR, and inserted into the pShuttle-CMV vector to construct the recombinant plasmids which were linearized by Pme I and transformed into BJ5183 cells containing the pAdEasy-1 bone plasmid. The infective replication-defective adenovirus rAd-3C was collected after the appearance of CPE, and it was used to infect Vero cell with rAd-P1, rAd-P1-2A, rAd-L-P1 and rAd-L-2A, respectively. It was showed that P1 protein and L-P1 protein were failed to be cleaved by 3C proteinase, while P1 protein was cleaved by 3C proteinase and 2A proteinase. The results of indirect ELISA showed that a certain level of antibodies were induced in mice immunized by the recombinant virus. These results would facilitate to further studies on the recombinant adenovirus live vector vaccine against FMD.

关 键 词：口蹄疫病毒 重组腺病毒 免疫原性 

分 类 号：S852.65[农业科学—基础兽医学]