茶毛虫感染核型多角体病毒(EpNPV)的快速分子鉴定

Rapid Molecular Identification of Euproctis pseudoconspersa Infected with EpNPV by PCR Method

根据茶毛虫核型多角体病毒(Euproctis pseudoconspera nuclear polyhedrosis virus,EpNPV)两个核心基因A13-1lef-8和A13-1polyhedrin的核苷酸序列,设计合成了两对特异性引物,应用PCR方法分别扩增获得538bp和281bp的2个DNA片段。克隆至T载体测序后与Genbank中已知EpNPV两基因序列比对,匹配率为100%。证实所克隆的特异性DNA片段可以作为鉴定茶毛虫感染EpNPV的标志性序列。本研究建立了茶毛虫感染EpNPV的分子鉴定方法,并为EpNPV防治茶毛虫效果的评价以及EpNPV侵染茶毛虫途径等毒理学研究提供依据。

According to nucleotide sequences of the two key genes, A13-1 lef-8 gene and A13-1 polyhedrin gene of EpNPV, two pairs of special primers were designed. Two typical DNA fragments with the size of 538 bp and 281 bp were amplified by PCR method. The two PCR products were cloned into T vector and sequenced, Blasted with the DNA sequence of EpNPV, both of them were matched absolutely. That meant that these two DNA fragments can be used as molecule markers to identify the Euproctis pseudoconspersa Strand infected with EpNPV. The present investigation constructed a molecular biological method to detect EpNPV DNA, to evaluate the biological control effect of EpNPV and to study the infecting route of EpNPV into E. pseudoconspera.