马铃薯Y 病毒河北分离物外壳蛋白基因序列分析和株系鉴定

 对河北张家口感病马铃薯的一个马铃薯Y病毒分离物( PXY-HBEI) 的外壳蛋白基因进行了克隆和序列分析。以提取的RNA为模板, 应用RT-PCR扩增目的基因, 扩增产物克隆到质粒pGEM-T, 上并序列分析。结果表明, 克隆cDNA片段由801个核苷酸组成, 编码267个氨基酸。与基因库中PVY代表株系相比, 它们的核苷酸和氨基酸序列同源性分别为89.6%～98.8%和93.3% ～98.5% , 与PVYN 和PVYO株系的氨基酸序列同源性分别为93.6%和97.8% , 确定PVY-HBEI归属于普通株系( PVYO株系) , 同时建立了快速、灵敏、简便的PVY RT-PCR检测方法。

Coat Protein Gene Sequence Analysis and Identification of a Potato Virus Y Hebei Isolate

A potato virus Y isolate ( PVY-HBEI) was obtained from infected potato plants cultivated in Zhangjiakou City of Hebei Province. The coat protein gene (CP) was amplified from the extracted plant total RNA by using RT-PCR, and cloned into the pGEM2T vector and sequenced. The cloned segmentwas 801 bp and encodes 267 amino acid residues. It was cpmpared with other PVY isolates reported in GenBank. It shares 89.6% - 98.8% and 93.3% - 98.5% homology in nucleotide and the putative amino acid sequence, respectively, which suggested that the CP gene of PVY isolates is a relatively conserved sequence within the PVY genome, the homology between PVY-HBEI and PVYN and PVYO were 93.6% and 97.8% , resepectivly, thus itwas identified as an isolate of PVYO , the experiment also provided a rapid, sensitive and relatively inexpensive methods of PVY RT - PCR detection.