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为研究马铃薯Y病毒的检测方法,参考GenBank中马铃薯Y病毒的全基因序列,应用Primer 6.0引物软件设计并合成了一对特异性引物PVYF、PVYR,利用RT-PCR方法对PVY病毒黑龙江分离株CP基因进行特异性扩增,对扩增片段进行序列测定并进行序列比对。结果表明:引物PVYF、PVYR可以扩增出包含PVY病毒CP基因全长的cDNA特异性片段,序列分析结果表明PVY病毒黑龙江分离株CP基因与GenBank上的40个不同PVY病毒分离株的CP基因氨基酸序列同源性为92.1%~97.4%,说明PVY病毒CP基因保守性较高,可以用作制备PVY病毒血清型检测方法的抗体基因。
In order to study the detection method of potato virus Y, a pair of specific primers PVYF and PVYR were designed and synthesized by using Primer 6.0 primer software with reference to the complete genome sequence of potato virus Y in GenBank. By using RT-PCR method, The genes were specifically amplified, the amplified fragments were sequenced and the sequences were aligned. The results showed that the primers PVYF and PVYR could amplify the cDNA fragment containing the full length of CP gene of PVY virus. The sequence analysis showed that CP gene of PVY virus isolated from Heilongjiang isolates of PVY virus was identical with CP gene of 40 different PVY isolates in GenBank Sequence homology was 92.1% ~ 97.4%, indicating that CP gene of PVY virus is highly conserved and can be used as an antibody gene for the preparation of the PVY virus serotype detection method.