【摘要】：為了對(duì)豬繁殖與呼吸綜合征病毒(PRRSV)Nsp9基因的功能進(jìn)行預(yù)測(cè),利用RT-PCR法從細(xì)胞毒中擴(kuò)增PRRSV FS株的Nsp9基因,并將其克隆至pMD18-T載體上,測(cè)序得到Nsp9基因序列,利用多種生物信息學(xué)軟件分析Nsp9序列生物學(xué)信息。結(jié)果顯示,Nsp9基因全長(zhǎng)1 929bp,目的基因的蛋白分子質(zhì)量為70.5ku,pI為7.78,表明該蛋白不穩(wěn)定;抗原性分析顯示,Nsp9基因存在多個(gè)抗原位點(diǎn),柔韌性較好,多處位點(diǎn)的親水性較強(qiáng),適合作為單抗制備的表位;同種亞型不同毒株的Nsp9基因氨基酸同源性為96.9%~98.9%,這表明Nsp9基因高度保守,可進(jìn)一步作為檢測(cè)靶蛋白用于豬繁殖與呼吸綜合征(PRRS)的診斷與疫苗免疫;亞細(xì)胞定位預(yù)測(cè)該基因可能定位于細(xì)胞質(zhì)中,而不是細(xì)胞核內(nèi);同源建模預(yù)測(cè)三級(jí)結(jié)構(gòu)結(jié)果顯示,三維結(jié)構(gòu)呈右手型,具有3個(gè)亞結(jié)構(gòu)域,沒(méi)有信號(hào)肽。此外,親本株Nsp9基因與疫苗株Nsp9基因存在多個(gè)氨基酸的突變,這些氨基酸的插入與缺失是否與病毒的聚合酶活性和毒力有關(guān)還需要進(jìn)一步試驗(yàn)驗(yàn)證。
[Abstract]:In order to predict the function of (PRRSV) Nsp9 gene of pig reproductive and respiratory syndrome virus, the Nsp9 gene of PRRSV FS strain was amplified from cytotoxicity by RT-PCR and cloned into pMD18-T vector, and the sequence of Nsp9 gene was obtained by sequencing. The biological information of Nsp9 sequence was analyzed by using a variety of bioinformatics software. The results showed that the full length of Nsp9 gene was 1929bp. the protein molecular weight of the target gene was 70.5kuand Pi was 7.78, indicating that the protein was unstable. Antigenicity analysis showed that Nsp9 gene had multiple antigen sites, good flexibility and strong hydrophilicity, so it was suitable for monoclonal antibody preparation. The amino acid homology of Nsp9 gene of different subtypes of the same subtypes was 96.9% 鈮，

本文編號(hào)：2478486