番木瓜eIF4E突变基因的构建

真核翻译起始因子4E（eukaryotic translation initiation factor4E，eIF4E）的缺失或突变能影响病毒对植物的侵染，并介导植物对病毒产生抗性。已有研究证明，番木瓜环班病毒的VPg能与番木瓜elF4E（Cp－eIF4E）互作，在此基础上，笔者通过蛋白序列比对和同源建模分析，确定了6个突变位点，采用套叠PCR方法印Cp－eIF4E基因进行点突变，然后，将它们分别连接到酵母双杂交系统和双分子荧光互补系统两套表达载体上，为后续互作实验和抗病功能验证奠定基础。

The Construction of Mutant eIF4E in Papaya

Mutations in or loss of the eukaryotic translation initiation factor 4E （eIF4E） can affect infections of viruses to plants, which leads to resistance of the plants against the viruses. The interaction between PRSV-Vpg and Cp-eIF4E（eIF4E in Papaya） has been verified. In this study, six point mutations were determined by protein sequence alignment and analysis of homology modeling, and the point mutations were produced in Cp-eIF4E by overlap extension PCR. Then, the mutant Cp-eIF4E genes were connected into two sets of expressive vectors, yeast two-hybrid system and bimolecular fluorescence complementation, which lays a foundation for the follow-up verification of interaction and resistance in the plants.