【摘要】 为了建立以本氏烟为模式作物研究SMV侵染机制的平台,以可系统侵染本氏烟的重组型SMV分离物HBRS为毒源,构建感染HB-RS本氏烟的酵母双杂c DNA文库。利用SMART技术获得本氏烟的双链c DNA文库,通过双酶切手段将cDNA文库构建至pGADT7-SfiI载体上。检测结果显示,文库容量约为3.2×106cfu,插入cDNA片段长度为0.75~2.00 kb,平均长度大于1.00 kb,且多态性丰富,表明文库质量较好。该酵母双杂文库的构建为钓取与SMV互作的寄主因子和系统性研究SMV侵染机制奠定了物质基础。更多还原

【Abstract】 In order to establish a platform for studying on the mechanism of SMV infection on the model crop of Nicotiana benthamiana. In this study,a recombinant SMV isolate HB-RS,which could systematically infect Nicotiana benthamiana,was used as virus source to construct a yeast two hybrid cDNA library of Nicotiana benthamiana with HB-RS infection. Double stranded cDNA library of the Nicotiana benthamiana were obtained through SMART technology and cDNA library were cloned into p GADT7-SfiI vector by double enzyme digestion method. Testing result showed that the cDNA library contained more than 3. 2 × 106 cfu and the cDNA fragments length inserted were from 0. 75- 2. 00 kb. The fragment was polymorphism and average length was more than 1. 00 kb indicating that the cDNA library was high quality. In all,cDNA library of the yeast two hybrids provided a material basis for screening host factor with SMV proteins and systematic studying of SMV infection mechanism.更多还原