甜菜夜蛾核多角体病毒Se29基因的克隆与序列分析

摘要

[目的]克隆得到甜菜夜蛾核多角体病毒(Spodoptera exigua muhicapsid 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 armigera single nueleocapsid nucleopolyhedrovir-as,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. NC_002169) of Se29. Predicted protein from nucleotide sequence of Se29 ( SE29 ) had one notable transmembrane region, and contained much PKC - PHOSPHO - SITE, CK2 -PHOSPHO - SITE ,ASN_GLYCOSYLATION. 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.