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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p BI121重组植物表达载体p BI121-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。
Objective To identify the virus isolates from Chuxiong tomato infected materials in Yunnan Province and establish the plant N expression vector for resistance breeding. Methods The N gene was cloned by RT-PCR and was 777 bp in length. The cloned TSWV-N gene was ligated into the plasmid pBI121 recombinant plant expression vector pBI121-TSWV-N. Results The TSWV-N gene shared 97% -100% identity with the registered tomato spotted wilt virus Yunnan isolate in China. Conclusion The isolated virus is indeed tomato spot wilt virus. The plant expression vector containing TSWV-N gene was successfully constructed.