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.
出处
《生物工程学报》
CAS
CSCD
北大核心
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。~~
