甜瓜抗枯萎病基因表达载体的构建及其转化  

Construction of Expression Vector with R-Fom-2 Gene and Its Genetic Transformation in Melon

作　　者：颜雪[1] 赵惠新[1] 王贤磊[1] 康锋[1] 李冠[1] 

机构地区：[1]新疆大学生命与科学技术学院,新疆乌鲁木齐830046

出　　处：《生物技术》2008年第3期9-11,共3页Biotechnology

基　　金：国家自然科学基金项目资助(30660078);自治区高校重点项目资助(xjedu2004I09)

摘　　要：目的:构建甜瓜抗枯萎病基因的植物表达载体,并将其转入新疆感病甜瓜。方法:含有目的片段的质粒pMD18-T/R-Fom-2经SacⅠ和SalⅠ双酶切后,回收目的片断,将该片断连接到pCAMBIA1301-1上,得到R-Fom-2基因的植物表达载体pCA-Fom-2。通过冻融法将该载体导入根癌农杆菌LBA4404菌株中,并转化新疆感病甜瓜。结果:获得了具有潮霉素抗性的再生植株,经过PCR和RT-PCR鉴定结果表明,目的基因已经整合到甜瓜基因组中并初步表达。结论:通过根癌农杆菌介导法将外源基因成功导入甜瓜中。The expression vector of R - Fore - 2 gene was constructed and transformed into radon to get the transgenic plants.Method： The cloned vector pMD18 - T/R - Fom - 2 and expression vector pCAMBIA1301 - 1 were digested with restriction nuclease Sac I and Sol I. The large fragments were purified. The two large fragments of pMD18 - T/R - Fom - 2 and pCAMBIA1301 - 1 were ligated by T4 DNA ligase and the reactant was transferred into E. coli. The recombinant binary vector was transfered into Agrobacterium and infected melon. Result： After selection with antibiotic,the green plant showed positive for RT- PCR.The fragment of aimed gene was purified after enzymic digestion plasmid pMD18 - T/R- Fom - 2 with Sac I and Sal I and cloned into pCAMBIA1301 - 1 to obtain the recombinant plant expression vector pCA - Fom - 2.

关 键 词：甜瓜 R-Fom-2 载体构建 遗传转化 

分 类 号：Q782[生物学—分子生物学] Q785