中国动物传染病学报2017，Vol.25Issue(2)：49-55,7.
新型鸭细小病毒NS1基因的原核表达及生物信息学分析
PROKARYOTIC EXPRESSION AND BIOINFORMATICS ANALYSIS OF NS1 GENE OF NOVEL DUCK PAROVIRUS
摘要
Abstract
In order to investigate the biological function of NS1 protein of Nocel duck parvovirus (NDPV), one pair of primers were designed according to the NS1 gene sequence of NDPV DS15 strain (GenBank No. KU947420). The PCR products from the NDPV NS1 gene were cloned into the prokaryotic expression vector pGEX-4T-1, resulting in the recombinant plasmid pGEX-4T-NS1. After sequencing analysis, bioinformatics software and net servers were performed to predict and analyze the structures and biochemical properties of NS1 protein. The recombinant plasmid was transformed into BL21 competent cells and then induced with IPTG. Expression of the NS1 protein was examined in SDS-PAGE and Western blot. The results showed that the recombinant protein NS1 with molecular weight of 97 kDa was expressed successfully in the prokaryotes. Bioinformatic analysis showed that the NS1 protein, a soluble and acidic protein, was mainly expressed in the cytoplasm and composed of 627 amino acids with a molecular weight of about 72 kDa. It contained one serine/threonine kinase substrate motif and one s546 potential phosphorylation site but had neither signal peptide nor transmembrane region. The main epitope mainly located at the C terminal end of NS1 polypeptide. Phylogenetic tree and homological analysis of NS1 gene showed that NDPV and goose Parvovirus (GPV) were grouped in the same evolutionary branch, which indicated that they had the closest genetic evolutionary relationship. The results of the present study have laid a foundation for revealing the role of the NS1 protein in the replication, proliferation and pathogenesis of NDPV.关键词
新型鸭细小病毒/克隆/原核表达/生物信息学Key words
Novel duck parvovirus/cloning/prokaryotic expression/bioinformatics分类
农业科技引用本文复制引用
张经伟,李琦,陈宗艳,宫晓华,李国新,温建新,朱杰,李传峰,刘光清..新型鸭细小病毒NS1基因的原核表达及生物信息学分析[J].中国动物传染病学报,2017,25(2):49-55,7.基金项目
国家自然科学青年基金 (31502068) （31502068）
上海市科委科技创新医药与农业领域项目(13391901602) （13391901602）
