番茄斑萎病毒N基因克隆及其植物表达载体的构建

Cloning of N gene of Tomato spotted wilt virus and construction of its plant expression vectors

中文关键词:  番茄斑萎病毒  N基因  植物表达载体

英文关键词:Tomato spotted wilt virus  N gene  plant expression vector

中文摘要:

      目的 对从云南楚雄番茄染病材料上得到病毒分离物进行鉴定, 建立病毒N基因植物表达载体, 以便进行抗病育种研究。方法 利用RT-PCR技术克隆得到N基因, 全长777 bp, 将克隆得到的TSWV-N基因连接至质粒pBI121重组植物表达载体pBI121-TSWV-N。结果 TSWV-N基因与NCBI上已登记的番茄斑萎病毒中国云南分离物同源性达到97%~100%。结论 所分离病毒确为番茄斑萎病毒, 包含有TSWV-N基因的植物表达载体构建成功。

英文摘要:

      Objective To detect and identify the virus from infected tomato plants in Chuxiong, Yunnan province, and construct the recombinant plasmid with the gene N for expressing in plant. Methods The nucleotide sequences of nucleocapsid protein (N) gene of Tomato spotted wilt virus (TSWV) were determined by RT-PCR with the length of 777 bp. Then the cloned TSWV-N gene was connected to pBI121-TSWV-N, which was the recombinant plasmid plant expression vector of pBI121. Results Compared with sequences of corresponding viruses from Yunnan in China that were available in the GeneBank, the N gene of TSWV-Chuxiong shared similarity of 97%~100% at nucleotide level, were ligated into the plant expression vectors, PBI121. Conclusion The virus was identified as TSWV, and the recombinant plasmid was constructed successfully by restriction enzyme analysis.