甜菜夜蛾核多角体病毒VP39基因的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of Spodoptera Exigua Multicapsid Nucleopolyhedrovirus VP39 Gene

作　　者：李赛男[1] 李萌 赵海洲[1] 姜玉霞[1] LI Sainan;LI Meng;ZHAO Haizhou;JIANG Yuxia(Biology Department,Zhaoqing University,Zhaoqing 526061,China)

机构地区：[1]肇庆学院生命科学学院

出　　处：《重庆理工大学学报（自然科学）》2019年第8期190-197,共8页Journal of Chongqing University of Technology：Natural Science

基　　金：国家自然科学基金资助项目(31501705);广东省自然科学基金资助项目(2014A030313664)

摘　　要：VP39是杆状病毒核衣壳主要结构蛋白基因,为研究VP39在杆状病毒生活周期中的可能作用,用PCR方法克隆得到甜菜夜蛾核多角体病毒(Spodoptera exigua multicapsid nucleopolyhedrovirus,SeMNPV)VP39全长基因(Sevp39),将其克隆至pMD18-T载体上,获得重组质粒T-Sevp39,进行序列测定和生物信息学分析。序列测定结果表明:扩增序列大小为981 bp,与GenBank登录序列(Genbank accession No.NC_002169)核苷酸同源性99.9%,第308位的碱基由G突变成T。序列分析表明:Sevp39编码蛋白(Sevp39)有多个可能的N-糖基化位点、酪蛋白激酶Ⅱ磷酸化位点和蛋白激酶C磷酸化位点,1个酪氨酸激酶磷酸化位点和1个N-豆蔻酰化位点。蛋白序列比对结果表明:Sevp39与绝大部分杆状病毒VP39的序列相似性均大于30%,在Sevp39同源蛋白中有6个高度保守的半胱氨酸残基(Cys)。蛋白质二级结构预测结果显示:Sevp39可能形成7个α螺旋和14个β折叠,高度保守的Cys^17参与了β折叠的形成。上述结果为研究Sevp39在病毒入侵和病毒复制过程中所担负的功能奠定了实验基础。VP39 is the main structural protein gene of baculovirus nucleocapsid.To study the possible role of VP39 in the baculovirus life cycle,the ORF of VP39 gene(Sevp39)was obtained by PCR method fromSpodoptera exigua multicapsid nucleopolyhedrovirus(SeMNPV)genomic DNA.The PCR product was cloned into pMD18-T vector to get the recombinant plasmid(T-Sevp39).Then Sevp39 gene was sequenced and analyzed.Nucleotide sequence analysis showed that the amplified sequence was 981 bp in length,shared 99.9%identity with the sequence of GenBank(Genbank accession No.NC_002169)of Sevp39.The predicted protein from nucleotide sequence of Sevp39(Sevp39)contained much PKC-PHOSPHO-SITE,ASN-GLYCOSYLATION,CK2-PHOSPHO-SITE.BLAST results showed that the amino acid sequence identity between Sevp39 and most baculovirus VP39 was greater than 30%.The amino acid sequence alignment of Sevp39 homologs showed that there were 6 highly conserved cysteine residues(Cys)in the Sevp39 homologs.The prediction of secondary structure showed that Sevp39 may form 7 helixes and 14 strands,and the highly conserved Cys^17 was involved in the formation of strand.These achievements lay an experimental foundation for further research of the gene functions in the course of virus invasion and replication.

关 键 词：甜菜夜蛾核多角体病毒 Sevp39 克隆 生物信息学分析 

分 类 号：Q785[生物学—分子生物学]