甜菜夜蛾核多角体病毒Se29基因的克隆与序列分析  

Cloning and Sequence Analysis of Spodoptera Exigua Multicapsid Nucleopolyhedrovirus Se29 Gene

作　　者：李赛男[1] 李充璧[1] 龚敬文[1] 陈焕楷[1] 

机构地区：[1]肇庆学院生命科学学院,广东肇庆526061

出　　处：《安徽农业科学》2011年第14期8520-8523,共4页Journal of Anhui Agricultural Sciences

基　　金：广东省教育厅自然科学基金重点项目(06Z026);肇庆市科技创新项目(203301);中山大学生物防治国家重点实验室开放课题(2004)

摘　　要：[目的]克隆得到甜菜夜蛾核多角体病毒(Spodoptera exiguamulticapsid nucleopolyhedrovirus,SeMNPV)ORF29全长基因(Se29)。[方法]用PCR的方法扩增Se29基因,将其克隆至pMD18-T载体上,获得了重组质粒T-Se29,进行序列测定和分析。[结果]获得大小为642 bp左右的序列,扩增序列与GenBank登录序列(Genbank accession No.NC_002169)核苷酸同源性100%。序列分析结果表明,Se29基因编码蛋白(SE29)存在一个可能的跨膜区域,有多个蛋白激酶C、酪蛋白激酶Ⅱ磷酸化位点和N-糖基化位点,1个微体C末端靶信号位点。蛋白序列比对结果表明,SE29与棉铃虫单核衣壳核多角体病毒(Helicoverpa armigerasingle nucleocapsid nucleopolyhedrovir-us,HaSNPV)ORF128(Ha128)的编码蛋白(HA128)具有较高的同源性,其在病毒侵染过程中的作用值得关注。[结论]成功克隆出Se29基因,为研究其在病毒侵染过程中的作用奠定了试验基础。[Objective] The aim was to obtain Se29 gene from Spodoptera exigua multicapsid nucleopolyhedrovirus（SeMNPV） genomic DNA.[Methods] The sequence of Se29 gene was obtained by using PCR method.The PCR product was cloned into pMD18-T vector to get the recombinant plasmid（T-Se29）.Then Se29 gene was sequenced and analyzed.[Result] Nucleotide sequence analysis showed that the amplified sequence was 642 bp in length,shared 100 % identity with the sequence of GenBank（Genbank accession No.NC002169） of Se29.Predicted protein from nucleotide sequence of Se29（SE29） had one notable transmembrane region,and contained much PKC-PHOSPHO-SITE,CK2-PHOSPHO-SITE,ASNGLYCOSYLATION.BLAST protein results showed that SE29 had high homology with HA128 protein encoded by Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus（HaSNPV） ORF128（25%）.The function of SE29 in the course of virus infection claims attention.[Conclusion] In this study,Se29 gene was cloned.This lays a foundation for further research of the gene functions in the course of virus infection.

关 键 词：甜菜夜蛾核多角体病毒 Se29 克隆 序列分析 

分 类 号：S435.663[农业科学—农业昆虫与害虫防治]