柞蚕NPV多角体膜蛋白类似基因的克隆和序列分析

为探明柞蚕核型多角体病毒(Anthraea pernyi nucleopolyhedrovirus,ApNPV)基因组信息,从柞蚕尸体中分离纯化ApNPV,提取其基因组DNA,分别建立ApNPV DNA的HindⅢ和SalⅠ酶切片段文库。对插入片段进行测序,经过同源性分析发现,在2．9kb的SalⅠ片段上包含1个多角体膜蛋白类似基因,其读码框编码289个氨基酸,是一个晚期表达基因。氨基酸同源性比较的结果表明,ApNPV的多角体膜蛋白基因与云杉卷叶蛾(Choristioneure fumiferana)NPV的同源性较高,达91．7%,而与家蚕(Bombyx mori)NPV及苜蓿尺蠖(Autographa califorlica)MNPV的同源性较低,分别为65．1%和63．0%。该结果说明ApNPV与CfNPV在进化关系上较近,而与BmNPV和AcNPV在进化关系上较远。比较多种核型多角体病毒的多角体膜蛋白和多角体蛋白的氨基酸序列发现,这两种多角体结构蛋白的同源性具有相似的变化趋势。

Cloning and Sequence Analysis of the Antheraea pernyi Nucleopolyhedrovirus Polyhedral Envelope Protein Gene

In ovder to probe into the genomic information of Anthraea pernyi nucleopolyhedrovirus(ApNPV), the polyhedron of ApNPV were purified from larval cadaver of Anthraea pernyi and the genomic DNA of ApNPV was extracted,then digested by HindⅢand SalⅠ,respectively.The fragments after digestion were inserted into vector pUC18.300 positive clones were obtained and sequenced.The result of homological analysis showed that an open reading frame,which contains 289 amino acids,is more similar to the Choristioneure fu- miferana nucleopolyhedrovirus(CfNPV)pp34 gene(91.7%)than to that of Bombyx mori nucleopolyhedrovir- us(65.1%)and Autographa califorlica nucleopolyhedrovirus(63%).This result indicated that the relationship of ApNPV and CfNPV is closer than that of ApNPV and BmNPV or AcNPV.Amino acid identity of 15 polyhedral envelope protein genes and polyhedrin genes was compared,and its trend was similar.