葡萄扇叶病毒杭州分离物(GFLV-H)P_(38)蛋白基因克隆及序列分析

Cloning and sequence analysis of P_(38) gene of Grapevine fanleaf virus Hangzhou isolated.

为了证实GFLV-HP38蛋白的性质及其在寄主扩散中的作用,根据葡萄扇叶病毒(Grapevinefanleafvirus,GFLV)法国分离物F13RNA2序列合成特异性引物,利用RT-PCR从GFLV-H基因组中扩增出P38蛋白基因的cDNA片段,并克隆到pGEM-T-easyvector上。序列分析表明该基因包含1044个核苷酸,编码347个氨基酸。序列比较发现GFLV-HP38蛋白基因与GFLV-F13对应基因核苷酸和氨基酸的同源性分别为90%和96%;与线虫传多面体病毒属中的南芥菜花叶病毒(Arabismosaicvirus,ArMV)和番茄环斑病毒(Tomatoringspotvirus,TomRSV)对应基因氨基酸同源性分别为85%和38.6%,与烟草黑环病毒(Tabaccoblackringvirus,TBLV)、葡萄铬色花叶病毒(Grapevinechromemosaicvirus,GCMV)和树莓环斑病毒(Raspberryringspotvirus,RRSV)的同源性均低于30%。

To confirm the characteristics and function of P38 in the viral particles movement, the cDNA fragment of P38 gene of Hangzhou isolate of Grapevine fanleaf virus (GFLVH) was amplified by RTPCR with specific primers to the RNA2 sequence of GFLVF13 and was then inserted into pGEMTeasy vector. The result of sequence analysis indicated that this gene contained 1044 nucleotides encoding 347 amino acids and were 90% and 96% identical to GFLVF13 at nucleotide and amino acid level, respectively. Multiple sequence alignment of amino acid sequence showed that homology between P38 gene of GFLVH and that of Arabis mosaic virus (ArMV) and that of Tomato ringspot virus (TomRSV) was 85% and 38.6%, respectively, while its homology with Tabacco black ring virus (TBLV), Grapevine chrome mosaic virus (GCMV) and Raspberry ringspot virus (RRSV) was below 30%.