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大麦黄矮病毒GPV株系是我国特有的一个株系,与国外已报道的大麦黄矮病毒MAV,PAV,SGV,RPV和RMY在血清学上无反应.通过对GPV.外壳蛋白基因的序列分析,明确了病毒的外壳蛋白基因由603个核苷酸组成,编码201个氨基酸,分子量为22218ku.同MAV,PAV,RPV株系一样,在GPV外壳蛋白基因框架中(ORF)含有1个Vpg框架结构,分子量为17024ku.外壳蛋白基因序列同源性比较结果显示,GPV株系和RPV株系同源性较高,核苷酸和氨基酸的同源性分别为83.7%和77.5%.而与PAV和MAV株系同源性较低,核苷酸和氨基酸的同源性分别为56.9%,53.2%和44.1%,43.8%.按照GPV外壳蛋白基因序列,设计寡聚核苷酸引物,通过RT-PCR反应,得到GPV外壳蛋白基因全长cDNA,克隆到表达质粒,构建了高等植物表达载体pPPI1,pPPI2和pPPI5.
The barley yellow dwarf virus GPV strain is a unique strain in our country, which has no serological reaction with foreign reported barley yellow dwarf virus MAV, PAV, SGV, RPV and RMY.By sequence analysis of GPV coat protein gene , Which made it clear that the coat protein gene of the virus consists of 603 nucleotides and encodes 201 amino acids with a molecular mass of 22218 ku. Like the MAV, PAV and RPV strains, it contains one Vpg framework in the GPV coat protein gene frame (ORF) Structure and molecular weight of 17024ku.The comparison of the sequence homology of coat protein gene showed that the homology of GPV strain and RPV strain was high and the nucleotide and amino acid homologies were 83.7% and 77.5% The homology of nucleotide and amino acid was 56.9%, 53.2% and 44.1%, 43.8%, respectively, compared with that of MAV strain.According to the GPV coat protein gene sequence, oligonucleotide primers were designed and sequenced by RT -PCR. The full-length cDNA of GPV coat protein gene was obtained and cloned into the expression plasmid. The higher plant expression vectors pPPI1, pPPI2 and pPPI5 were constructed.