花生条纹病毒(红安分离物)cp基因的克隆和序列分析

Cloning and sequencing the coat protein gene of Peanut stripe virus Hongan isolate

从湖北省红安县花生种传苗中得到花生条纹病毒(Peanut stripe virus,PStV)分离物(PStV-Hongan),提取感病叶片的总RNA,RT-PCR扩增了其外壳蛋白基因(cp),克隆至pGEM-T Easy载体,转化大肠杆菌并鉴定。阳性质粒序列测定结果表明,该cp含有861个核苷酸,编码分子量为32.4kDa的蛋白。该株系cp与PStV其它株系核苷酸序列同源性为94.6%-98.8%,氨基酸序列同源性为96.2%-99.3%。株系间cp核苷酸序列系统进化树结果表明,Hongan株系与中国其它株系亲缘关系较近,而与东南亚其它株系关系较远。

Peanut stripe virus - Hongan isolate was obtained from the infected peanut seedling in Hongan County, Hubei Province. Its coat protein gene was amplified by RT - PCR and cloned into pGEM - T Easy vector. Positive clones were identified by PCR and restriction enzyme digestion. Sequence analysis of positive clone showed that the nucleotide sequence homology of PStV - Hongan shared 94.6 - 98.8% , amino acid sequence homology shared 96.2 - 99.3 % with other PStV strains. The strain shared closer relationship with Chinese strains compared with Southeast Asia strains by phylogenetic tree analysis of coat protein nucleotide sequence.