同安钮夜蛾核型多角体病毒pst I-G片段克隆及序列分析  

Sequence Analysis of the pst I-G Genomic Fragment of the Ophiusa Disjungens Nucleopolyhedrovirus

作　　者：常润磊[1] 林同[1,2] 张宇宏[1] 张琪[1] 温秀军[1] 

机构地区：[1]华南农业大学林学院,广州510642 [2]华南农业大学经济林研究中心,广州510642

出　　处：《中国农业科学》2009年第7期2373-2379,共7页Scientia Agricultura Sinica

基　　金：广东省自然科学基金(8151064201000068);华南农业大学经济林研究中心资助项目(4400-32990190);华南农业大学校长基金(4400-208012);华南农业大学新学科扶持基金(2008X039)

摘　　要：【目的】测定同安钮夜蛾核型多角体病毒(OpdiNPV)pst I-G片断的序列,了解OpdiNPV的遗传结构和与其它昆虫病毒的进化关系。【方法】用pstⅠ酶切病毒基因组,电泳分离并回收pst I-G片段,连接到PUC18,转化入E.coli DH 5α,挑取阳性菌落测序。【结果】该片段长度为5056bp,有4个ORF,分别编码ODV-E66的C末端(EU 623602),P87/VP80的C末端(EU 732665),ODV-Ec43(EU 617337)和Ac108(EU 732666)。ac108基因和odv-ec43起始密码子上游都有1个晚期启动子结构域TAAG,odv-ec43基因起始密码子上游有2个早期转录起始元件CAGT;由odv-ec43推导的蛋白有2个跨膜螺旋、3个N-糖基化位点、1个N端酰基化位点、7个PKC磷酸化位点、1个酪氨酸激酶磷酸化位点和4个CKⅡ磷酸化位点;p87/vp80基因终止密码子下游有polyA信号。【结论】氨基酸序列同源分析和相似性分析显示Ac108、ODV-E66和P87/VP80与其它昆虫病毒差异较大,ODV-Ec43的保守性较高。[ Objective ] In order to investigate the genomic organization and phylogenetic status of OpdiNPV. [ Method] OpdiNPV DNA was digested with pst I and separated by electrophoresis. Apst I-G fragment was recovered, ligated into PUCI8 vector, and transformed into E. coli strain DH5α. Recombinant plasmids were purified from white bacterial colonies and sequenced. [ Result ] The pst I-G fragment was 5056bp in length, containing four open reading frames （ORFs） that encode odv-e66 gene （EU 623602）, p87/vp80 gene （EU 732665）, odv-ec43 gene （EU617337） and ac108 gene （EU 732666） respectively. A late promoter motif TAAG was found 12 nucleotides （nt） upstream of the start codon ATG ofac108 gene. Upstream of the start codon ATG ofodv-ec43 has two early transcription motifs CAGT and a late promoter motif TAAG. Two transmembrane helices, 3 N-glycosylation sites, 1 N-myristoylation site, 7 Protein kinase C phosphorylation sites, 1 Tyrosine kinase phosphorylation site, 4 Casein kinase Ⅱ phosphorylation sites were found in the protein deduced by odv-ec43.A polyadenylation signal site was found 89 nucleotides （nt） downstream of the stop codon TAAofp87/vp80 gene. [Conclusion] Amino acid sequence alignment and identity analysis showed that Ac 108, ODV-E66 and P87/VP80 had a low identity with other baculovirus. ODV-Ec43 was conserved among baculoviruses.

关 键 词：同安钮夜蛾核型多角体病毒 odv-e66基因 odv-ec43基因 ac108基因 p87/vp80基因 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]