为了探讨质型多角体病毒基因的遗传与变异,通过RT—PCR从家蚕(Bombyx mori)质型多角体病毒(Cytoplasmic Polyhedrosis Virus,CPV)苏州株(BmCPV—suzhou)的dsRNA中成功克隆了S10片段。该片段cDNA含有一个744bp的完整开放阅读框(ORF),编码由248个氨基酸残基组成的多角体蛋白,其分子量为28.46kD,等电点为7.12。比较不同宿主来源的CPV多角体蛋白基因氨基酸序列发现,同型CPV多角体蛋白基因高度相似,而不同型的CPV多角体蛋白之间几乎没有同源性;基于质型多角体蛋白基因的核苷酸序列和氨基酸序列的系统进化分析表明,BmCPV的不同株系与舞毒蛾(Lymantria dispar)CPV(LdCPV)和马尾松毛虫(Dendrolimas punctatus)CPV(DpCPV)聚为一类,表明三种CPV的亲缘关系较近。研究结果为深入探讨质型多角体病毒的变异与分子进化提供了新的参考。
In order to discuss the heredity and variation of Cytoplasmic polyhedrosis virus ( CPV), S10 fragment has been cloned successfully from dsRNA of Bombyx mori cytoplasmic polyhedrosis virus Strain Suzhou (BmCPV-suzhou) by RT-PCR. The S10 fragment contains a open reading frame (ORF) with 744bp encoding the polyhedrin, which is composed of 248 amino acid residues and its molecular weight is 28.46kD, the isoelectric point is 7. 12. Compared with amino acid sequences of the CPV polyhedrin genes from different hosts, the homeotypic CPV polyhedrin genes are highly similar, while no comparability among heterotypic CPV polyhedrin genes. Based upon amino acid and nucleotide sequences of the polyhcdrin, the phylogenetic tree of Cypovirus showed that BmCPVs, Lymantria dispar CPV and Dendrolimus punctatus CPV formed a cluster , indicating that the relationship is closer among them. The results provide a new reference for the further discussion on the mutation and molecular evolution of cytoplasmic polyhedrosis virus.