甘薯羽状斑驳病毒中国分离株外壳蛋白基因的克隆和序列分析

Cloning and Sequence Analysis of Coat Protein Gene of Sweet Potato Feathery Mottle Virus Chinese Isolate (SPFMV-Ch)

根据已报道的甘薯羽状斑驳病毒(Sweetpotatofeatherymottlevirus,SPFMV)外壳蛋白基因序列,设计合成了一对特异性引物,以我们从国内甘薯品种徐薯-18上分离到的SPFMV的RNA为模板,经过反转录合成cDNA第一条链,经PCR扩增、限制性酶切后克隆于pUC19的Hind 和SacI位点,转化大肠杆菌JM109,经限制性酶切分析、PCR鉴定以及序列分析证实获得了SPFMV中国分离株外壳蛋白基因的全长克隆.

The cDNA of SPFMV coat protein gene was synthesized by RT-PCRwith artifically synthesized two specific primers and the RNA of sweet potato feathery mottle virus Chinese isolate as templete. The synthesized cDNA was cloned into plasmid pUC19 in E. coli JM109. The recombinant plasmid was identified by PCR, restriction endonuclease and sequence analysis.The tests show that the coat protein gene of SPFMV-Ch was obtained.