【摘要】 应用分段RTPCR方法和分子克隆技术从家蚕质型多角体病毒(Bombyxmoricytoplasmicpolyhedrosisvirus,BmCPV)中国株的dsRNA中成功地分3段克隆了RDRP基因(RNAdependentRNApolymerase),大小分别为1442、827、1675bp,进行了基因全序列拼接,获得37kb的全长RDRP基因(GenBank序列号为AY496445)。通过在线blast分析,与BmCPV1、DpCPV1、LdCPV1、LdCPV14、TnCPV15核苷酸同源性分别为89%、81%、81%、541%和509%,氨基酸同源性分别为965%、931%、929%、411%和335%。根据核苷酸同源性与氨基酸同源性构建的进化树具有较好的一致性。通过ClustalX软件分析,定位了该病毒RDRP基因的3个保守区域(酸性区,核苷酸结合的核心区和催化功能核心区)。更多还原

【Abstract】 Step-by-step RT-PCR and molecular cloning technique were successfully applied to clone the three segments (1 442,827,1 675 bp) of the RDRP gene of the BmCPV(China strain).The whole RDRP gene of ~3.7 kb was formed and was sequenced by PCR and other methods.(GenBank Accession Number:AY496445).Identities of the nucleotide sequences between BmCPV-C and BmCPV-1,DpCPV-1,LdCPV-1,LdCPV-14,TnCPV-15 were 89%,81%,81%,54.1% and 50.9% respectively and that of the amino acid sequences were 96.5%,93.1%,92.9%,41.1% and 33.5% respectively.Phylogenetic trees created from the alignment of the nucleotide sequences and the alignment of amino acid sequences showed a good agreement.Three conserved regions in RDRPs(acid region,binding site of the nucleotides and catalytic core region) were located using software ClustalX.更多还原