口蹄疫病毒亚洲I型YNBS/58株P12A-3C转基因植物双元表达载体的构建  

Construction of the binary expression vector of transgenic plant with foot and mouth disease virus, Asia-1 type, YNBS/58 strain

作　　者：王炜[1] 张永光[1] 王永录[1] 方玉珍[1] 潘丽[1] 蒋守田[1] 刘力宽[1] 孙元[1] 吕建亮[1] 智晓莹[1] 黄振华[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：《中国人兽共患病学报》2006年第11期1059-1061,1064,共4页Chinese Journal of Zoonoses

基　　金：863国家高技术研究发展计划资助(2003AA241110)

摘　　要：目的双元载体是目前转基因植物工程普遍应用的载体,本文将口蹄疫病毒P12A基因、3C基因串联,构建植物表达载体pBin-438P12A-3C。方法为便于实验操作,本试验采用先将基因片段P12A、3C构建到一个中间质粒pcDNA3.1(+)的策略,构建成pcDNA3.1(+)-P12A-3C。结果通过对构建好的pBin438-P12A-3C(Mini-Ti)重组质粒进行PCR鉴定和序列测定,证明中间表达载体构建成功。结论通过三亲杂交法将重组质粒载体pBin438-P12A-3C导入到农杆菌GV3101,通过抗性筛选、PCR鉴定和序列测定,证明构建成功,与农杆菌中的help-Ti质粒共同组成植物双元表达载体,为以后的植物转化奠定基础。At present, the binary vectors are the most commonly used expression vector in the researches of transgerdc plant engineering. In the present study, the plant expression vector pBin-438P12A-3C was constructed using the connection of P12A gene and 3C gene in foot and mouth disease virus （FMDV）. For the sake of convenience of experimental works, the gene fragments of P12A and 3C were at first constructed into an intermediate plasmid pcDNA3.1 （＋）, and then the vector pcDNA 3. 1（＋）-P12A-3C was constructed and analyzed by PCR and sequencing. It was proved that the plant expression vector pBin- 438P12A-3C was successively constructed as demonstrated by PCR identification, sequence determination and resistance screening, and it could be transferred into Agrobacterium tumefaciens GV3101 by triparental mating.

关 键 词：口蹄疫病毒 转基因植物 表达载体 农杆菌 

分 类 号：R382.5[医药卫生—医学寄生虫学]