猪繁殖与呼吸综合征病毒FS株Nsp9基因的克隆与序列分析

发布时间：2019-05-16 18:44

【摘要】：为了对猪繁殖与呼吸综合征病毒(PRRSV)Nsp9基因的功能进行预测,利用RT-PCR法从细胞毒中扩增PRRSV FS株的Nsp9基因,并将其克隆至pMD18-T载体上,测序得到Nsp9基因序列,利用多种生物信息学软件分析Nsp9序列生物学信息。结果显示,Nsp9基因全长1 929bp,目的基因的蛋白分子质量为70.5ku,pI为7.78,表明该蛋白不稳定;抗原性分析显示,Nsp9基因存在多个抗原位点,柔韧性较好,多处位点的亲水性较强,适合作为单抗制备的表位;同种亚型不同毒株的Nsp9基因氨基酸同源性为96.9%~98.9%,这表明Nsp9基因高度保守,可进一步作为检测靶蛋白用于猪繁殖与呼吸综合征(PRRS)的诊断与疫苗免疫;亚细胞定位预测该基因可能定位于细胞质中,而不是细胞核内;同源建模预测三级结构结果显示,三维结构呈右手型,具有3个亚结构域,没有信号肽。此外,亲本株Nsp9基因与疫苗株Nsp9基因存在多个氨基酸的突变,这些氨基酸的插入与缺失是否与病毒的聚合酶活性和毒力有关还需要进一步试验验证。
[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，

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