马铃薯卷叶病毒外壳蛋白基因克隆转化及其转基因后代的表达

根据GenBank中的马铃薯卷叶病外壳蛋白基因(PLRV-CP)全序列,设计一对特异性引物,以马铃薯卷叶病毒RNA为模板,克隆了马铃薯卷叶病外壳蛋白基因,在pBI121的基础上构建了植物表达载体。利用PLO(Poly-L-Ornithine)将PLRV-CP基因导入到马铃薯加工型品种大西洋的原生质体中,获得了转基因后代。在5株转化后代中扩增出长度为610bp的目标DNA片断,说明导入的外源PLRV-CP基因已经整合到马铃薯基因组中。Southern blot分析结果进一步证明了PCR结果的正确性。RT-PCR结果表明,在3株转化后代叶片中具有阳性表达。马铃薯卷叶病毒接种实验结果表明,转基因植株比对照有明显的马铃薯卷叶病抗性。

Cloning and Transformation and Expression of Potato Leafroll Virus Coat Protein Gene in Potato

A pair of specific primer had been designed according to full length sequence of Potato leafroll virus coat protein gene(PLRV-CP) in the GenBank.Potato leafroll virus coat protein gene(PLRV-CP) had been cloned with the RNA of Potato leafroll virus as moplate,and plant expression vector had been constructed.PLRV-CP gene had been transformed to protoplate of the potato variety Daxiyang by the method of PLO(Poly-L-Ornithine) and transgenic plants.About 610bp interest DNA fragment had been amplification in 5 transgenic plants.This showed that exogenous PLRV-CP had been integrated to the genomy of potato.And the result of the Southern blot proved the correct of the PCR and the result of the RT-PCR further demonsticated that: there are positive expression in the 3 progenies of transgenic plants.And the potato leafroll virus inoculation experiment show that transgenic plants Resistant to the potato leafroll compared with control plants.