口蹄疫病毒VP1基因在转基因拟南芥种子中的表达及活性检测  

Expression and Active Detection of the Gene Encoding the FMDV Structural Protein VP1 in Transgenic Arabidopsis thaliana

作　　者：潘丽[1] 张永光[1] 王永录[1] 方玉珍[1] 蒋守田[1] 王宝琴[1] 王文秀[1] 王炜[1] 孙元[1] 吕建亮[1] 谢庆阁[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室,兰州730046

出　　处：《畜牧兽医学报》2006年第5期469-473,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金："863"国家高技术研究发展计划(2001AA2130712003AA241110)

摘　　要：构建了FMDV VP1基因的种子特异性表达载体p7SBin438/VP1,在根癌农杆菌GV3101的介导下,通过浸花法转化拟南芥花序,转基因拟南芥通过卡那霉素抗性筛选后,进行了目的基因PCR扩增和ELISA检测,对ELISA筛选的阳性植株进行Western blot检测,并对转基因拟南芥后代进行了目的基因PCR分析。结果表明,种子特异性表达载体p7SBin438/VP1构建正确,FMDV结构蛋白VP1编码基因已整合进拟南芥基因组并获得表达,表达蛋白具有免疫反应活性,转基因拟南芥后代分析表明VP1基因已经遗传给后代。本试验为将FMDV免疫基因向豆科植物种子中的转化提供了依据。The plant seed-specific expression vector p7SBin438/VP1 for VP1 gene of foot-andmouth disease virus O/China/99 strain was constructed. Mediated with Agrobacterium tumefaciens GV3101 harboring p7SBin438/VP1, VP1 gene was transferred into Arabidopsis thaliana via floral dip. After selecting by Kanamysin, a number of rcsistant lines were obtained. The presence of VP1 gene in the transformed Arabidopsis thaliana genome was confirmed by PCR. The active detection of expressed target protein in the pods and leaves of transgenic Arabidopsis thaliana was confirmed by sandwich-ELISA and Western blot. Genetic analysis of transgenic progeny was performed by PCR. Experimental results showed that the plant seed-specific expression vector was successfully constructed and the structural protein VP1 was expressed in pods at high-level. Western blot showed that recombinant protein expressed in pods had immunocompetence and could react with FMDV antisera. Progeny test showed that VP1 gene in the transgenic Arabidopsis thaliana was heritable stablely. These results may provide the experimental bases for further research on transformed leguminous plant.

关 键 词：口蹄疫病毒 VP1基因 种子特异性表达载体 转基因拟南芥 

分 类 号：Q78[生物学—分子生物学] S852.65[农业科学—基础兽医学]