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 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.MethodsThe nucleotide sequences of nucleocapsid protein (N) gene ofTomato 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.ResultsCompared with sequences of corresponding viruses from Yunnan in China that were available in the GeneBank, the N gene of TSWV-Chuxiong shared similarity of97%~100% at nucleotide level, were ligated into the plant expression vectors, PBI121.ConclusionThe virus was identified as TSWV, and the recombinant plasmid was constructed successfully by restriction enzyme analysis.
出处
《食品安全质量检测学报》
CAS
2014年第12期3939-3943,共5页
Journal of Food Safety and Quality
基金
山东省科技专项(2011SDH204)
国家质检总局科技计划项目(2012IK281
2013IK173
2009IK254)
山东出入境检验检疫局科技计划项目(SK201308)~~
关键词
番茄斑萎病毒
N基因
植物表达载体
Tomato spotted wilt virus
N gene
plant expression vector
