作　　者： ; ;

机构地区： 华南农业大学资源环境学院

出　　处： 《华中农业大学学报》 2005年第4期325-329,共5页

摘　　要： 利用PCR重叠延伸技术(genesplicingbyoverlapextension,SOE)将番木瓜环斑病毒(Papayaring-spotvirus,PRSV)Vb株系的外壳蛋白(coatprotein,CP)基因和Ys株系的复制酶(replicaseprotein,RP)基因构建成融合基因VY。同时对常规的SOE法进行了改进,并对改进的SOE的反应体系进行了优化,其中以Py-robestDNA聚合酶和60℃的退火温度为最佳。将融合基因VY构建在植物表达载体pCAMBIA2300上,测序结果表明,该融合基因的序列与设计相符。融合基因VY植物表达载体的构建,可为进一步获得对PRSV广谱抗性的转基因番木瓜奠定基础。 The fusion gene VY containing coat protein （CP） gene of v 0 strata ana repncase protein （RP） gene of Ys strain of Papaya ringspot virus （PRSV） were obtained with technique of splicing by overlap extension（SOE）. Technique of standard SOE was improved and the factors that influenced PCR amplification of standard SOE were screened. The results showed that Pyrobest DNA polymerase and 60℃ of annealing temperature were optimal for obtaining the fusion gene VY. The fusion gene VY was cloned into the plant expression vector of pCAMBIA2300 and was confirmed by DNA sequence analysis.It provided a reliable basis for development of transgenic papaya varieties with broader resistance to PRSV.

关 键 词： 番木瓜 番木瓜环斑病毒 融合基因 法

领　　域： [农业科学] [农业科学] [农业科学]