MDMV CP基因的克隆及其转基因玉米的研究  

Isolation of the Capsid Protein Gene of Maize Dwarf Mosaic Virus and its Transformation in Maize

作　　者：刘小红[1] 张红伟[1] 刘昕 刘欣洁[1] 谭振波[1] 荣廷昭[2] 

机构地区：[1]北京农林科学院北京农业生物技术研究中心,北京100089 [2]四川农业大学玉米研究所,雅安625014

出　　处：《生物工程学报》2005年第1期144-148,共5页Chinese Journal of Biotechnology

基　　金：北京市高技术室基金项目 (No .95 3 85 0 10 0 )资助。~~

摘　　要：用RT_PCR方法分离了玉米矮花叶病毒外壳蛋白基因 (MDMVCP) ,并且利用基因枪法将该基因导入玉米优良自交系18_5 99红、18_5 99白幼胚诱导的愈伤组织中。转化的愈伤组织在Bialaphos浓度 (PPT)为 8mg L、10mgL、5mg L的筛选压下经过 3次抗性筛选后 ,分别再生出可育植株 12株和 6株。PCR和Southern检测结果说明CP基因已整合到玉米自交系基因组中。对T1代转基因植株进行病毒人工接种试验 ,结果表明对照植株全部表现为感染玉米矮花叶病的典型症状 ,而转基因植株后代呈现不同程度的抗性。The MDMV(Maize Dwarf Mosaic Virus, MDMV) CP (Coat Protein, CP) gene was cloned by RT_PCR method and introduced into the embryonic calli derived from immature embryos of elite inbred 18_599hong and 18_599bai via particle bombardment. Bombarded calli were selected on selection medium containing 5～10 mg/L ( PPT ) Bialaphos. From resistant calli, 79 plantlets were regenerated. 18 of 79 were grown and harvested. The results of Southern blotting and PCR analysis demonstrated that MDMV CP have been integrated into the genome of the transgenic plants. PCR_positive progeny plants were artificially inoculated with MDMV strain B, and the average chlorosis of the functional leaves of each plant was investigated. The typical symptoms were observed from the leaves of the control inbreds. while, the presence of the MDMV CP gene provided resistance to inoculation with MDMV strain B.

关 键 词：玉米矮花叶病毒 外壳蛋白 遗传转化 人工接种 

分 类 号：S513[农业科学—作物学]