Abstract: In order to prokaryotically express the CD163 protein, one receptor of porcine reproductive and respiratory syndrome virus (PRRSV), we predicted the structure of CD163 using ScanProsite software, and fragmented it. CD163 fragments from primary alveolar macrophages (PAM) were amplified by RT-PCR using specific primers based on the known sequence from pig PRRSV(GenBank accession no. EU016226). Three fragments of CD163 were amplified and cloned into the prokaryotic expression vector pET-28a (+), the recombination proteins of CD163 fragments were expressed in E. coli cells. Western blot test verified that the expressed recombinant proteins had good antigen activities.This work provided the experimental basis for further study of the CD163 receptors role in the process of viral infection, and the interaction with other receptors.

Key words: PRRSV, CD163, clone, prokaryotic expression

摘要： 为原核表达猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus, PRRSV)受体CD163,本研究根据CD163基因序列(EU016226),通过生物信息学软件Scanprosite预测了CD163结构特征并进行分段。利用DNAstar软件设计合成3对特异引物,从猪肺泡巨噬细胞(porcine alveolar macrophages, PAM)中克隆CD163三个片段,经测序鉴定正确后,分别克隆于原核表达载体pET-28a(+),在IPTG诱导下,进行CD163重组蛋白的截短表达,经Western杂交检测证实表达的三段截短蛋白均具有良好的与抗原反应活性,从而为进一步研究CD163受体在病毒感染过程中的作用以及与其他受体之间的相互作用提供实验依据。

关键词: PRRSV, CD163, 克隆, 原核表达