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利用马铃薯卷叶病毒（ＰＬＲＶ）复制酶基因３′端长约６００ｂｐ的ｃＤＮＡ，用缺口平移方法进行３２Ｐ同位素标记，制备成ｃＤＮＡ探针，通过核酸斑点杂交，对提纯的马铃薯卷叶病毒（ＰＬＲＶ）、病毒ＲＮＡ、ＰＬＲＶ感染的马铃薯汁液，表达ＰＬＲＶＣＰ基因的马铃薯叶片及块茎芽进行了检测．结果表明，３２Ｐ标记的复制酶基因ｃＤＮＡ探针特异性强、灵敏度高．可测得提纯病毒的最低含量为４０８ｎｇ／ｍｌ，病毒ＲＮＡ最低量为２７．８ｐｇ／ｍｌ，未转ＣＰ基因接种ＰＬＲＶ的马铃薯叶片提取液的最高稀释度为１３７５．接种ＰＬＲＶ的转ＣＰ基因的抗性马铃薯块茎芽和叶片提取液的最高稀释度为０～１／１５．此探针和只转入病毒外壳蛋白基因，不接种ＰＬＲＶ的转基因马铃薯汁液不发生反应．表明，探针只与ＰＬＲＶ基因组ＲＮＡ特异反应，而不与外壳蛋白基因及其ｍＲＮＡ反应，从而为表达ＣＰ基因的转基因马铃薯中ＰＬＲＶ的检测和抗病性鉴定建立了一种可靠的灵敏的分子生物学技术．此项技术已用于转ＣＰ基因马铃薯Ｄｅｓｉｒｅ，Ｆａｖｏｒｉｔａ，乌盟６０１和虎头的抗病性鉴定
CDNA of about 600 bp was amplified from the 3 ’end of the gene encoding the potato leaf curl virus (PLRV) and nicked by 32P using a gap-shifting method. A cDNA probe was prepared. Nucleocapsid spot blot hybridization was used to purify the potato leafroll virus (PLRV) , Viral RNA, potato juice infected with PLRV, potato leaves and tuber buds expressing the PLRVCP gene. The results showed that the 32P-labeled replicase gene cDNA probe has high specificity and high sensitivity. The lowest detectable level of purified virus was 408 ng / ml, the lowest level of viral RNA was 27.8 pg / ml, and the highest dilution of potato leaf extract with no CP gene inoculated with PLRV was 1375. The highest dilution of potato tuber bud and leaf extract resistant to CP gene vaccinated with PLRV was 0-1 / 15. This probe does not react with transgenic potato juice that has been transformed only with the viral coat protein gene and without PLRV. Indicating that the probe specifically reacts with the PLRV genomic RNA and not with the coat protein gene and its mRNA to establish a reliable and sensitive molecular biology for the detection and disease resistance identification of PLRV in the transgenic potato expressing the CP gene Learn technology. This technique has been used to transfer the CP gene to Desire, Favorita, Wumeng 601 and Tiger head disease resistance identification