A/O型FMDV多抗原表位融合基因植物表达载体构建及转化柱花草的研究  

Construction of Plant Expression Vectors Fused Multi-antigen Epitope Genes of FMDV Type O/A and Genetic Transformation in Stylosanthes spp.

作　　者：肖旭倩[1,2] 胡正龙 沈文涛 言普[1] 王冬梅[4] 周鹏 XIAO Xu-qian;HU Zheng-long;SHEN Wen-tao;YAN Pu;WANG Dong-mei;ZHOU Peng(Key Laboratory of Tropical Crop Biotechnology,Ministry of Agriculture,Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Hainan Haikou 571101,China;Department of Biology and Environment,Wanbo Institute of Science&Technology,Anhui Hefei 230001,China;Hefei Guozheng Assets Management Company Limited,Anhui Hefei 230001,China;Guangxi Academy of Agricultural Sciences,Guangxi Nanning 530007,China)

机构地区：[1]中国热带农业科学院热带作物生物技术研究所农业部热带作物生物技术重点实验室,海南海口571101 [2]万博科技职业学院生物与环境系,安徽合肥230001 [3]合肥市国正资产经营有限公司,安徽合肥230001 [4]广西农业科学院,广西南宁530007

出　　处：《西南农业学报》2018年第8期1565-1570,共6页Southwest China Journal of Agricultural Sciences

基　　金：国家"973"计划(2007CB108903);中央级事业单位基本科研业务费资助

摘　　要：【目的】获得含有A/O型FMDV多抗原表位融合基因4种不同组合的转基因柱花草植株。【方法】用限制性内切酶Xba I和Sac I双酶切4个中间载体和质粒pBI121,回收连接后获得4个重组植物表达载体(pBI-xsB/xs T/xsBT/xsBTT)及相应农杆菌工程菌株,利用农杆菌介导法转化热研2号柱花草(Stylosanthes guianensis cv.ReyanⅡ)子叶叶盘。采用卡那霉素筛选,特异引物和转基因检测引物进行PCR及RT-PCR检测不同组合多抗原表位融合基因的转录情况。【结果】获得36株抗性转化株,经PCR检测有16株为阳性,RT-PCR检测后有11株为阳性,表明4种组合多抗原表位融合基因在转基因植株中获得转录。【结论】此研究为进一步揭示同型与异型FMDV之间多抗原表位基因不同融合方式的免疫原性和免疫效果奠定了研究基础。【Objective】4 different combinations of transgenic plants containing multi-antigen epitope genes of FMDV type A/O would be obtained.【Method】4 different recombinant plant expression vectors(pBI-xsB/xsT/xsBT/xsBTT)were constructed successfully by restriction endonuclease Xba I and Sac I digestions of 4 intermediate vectors and plasmid pBI121 and ligation,and cotyledon leaves of Stylosanthes guianensis cv.Reyan with Agrobacterium tumefaciens EHA105 carrying 4 recombinant plant expression vectors respectively were transformed.After Kanamycin resistant selection,the transformed plants by PCR and RT-PCR were analyzed,which confirmed whether all the fused multi-antigen epitope genes were transcribed.【Result】36 resistant plants by PCR and RT-PCR were analyzed,and 11 resistant plants were comfirmed to be positive,which showed that the fused multi-antigen epitope genes in transgenic plants were transcribed.【conclusion】This study laid a foundation for further studying the immunogenicity and immune effects of different fused multi-antigen epitope genes between homotypic and heterotypic FMDV.

关 键 词：蹄疫病毒 多抗原表位融合基因 农杆菌介导法 柱花草 遗传转化 

分 类 号：S858[农业科学—临床兽医学]