| 摘要: |
| 从猪肺泡巨噬细胞(PAM)中提取总RNA为模板,采用RT-PCR法获得猪繁殖与呼吸综合征病毒(PRRSV)的受体CD163全基因,将该基因克隆到真核表达载体pcDNA3.1/V5-HIS A上,构建出真核重组质粒pcDNA3.1-CD163,经酶切和DNA测序证明获得了CD163基因,其序列与GenBank报道序列比较,核苷酸同源性为99.37%。将测序正确的CD163基因在脂质体LipofectamineTM2000介导下转染PK-15细胞,通过间接免疫荧光(IFA)检测到了CD163在PK-15中的表达。将转染的细胞感染PRRSV后,经IFA检测转染CD163的细胞能够被PRRSV感染。 |
| 关键词: 猪肺泡巨噬细胞 猪繁殖与呼吸综合征病毒 清道夫受体；CD163基因 |
| DOI： |
| 投稿时间：2013-06-29修订日期：2013-08-20 |
| 基金项目: |
|
| The Eukaryotic Expression and Functional Identification of Scavenger Receptor CD163 of PRRSV |
|
| (Animal Huabandry and Veterinary Research Institute,Shanghai Academy of Agricultural Science) |
| Abstract: |
| Total RNA was extracted from porcine alveolar macrophage, and then the recetor CD163 cDNA was obtained by RT-PCR. The full length CD163 was cloned into pcDNA3.1/V5-HIS A vector, and thus the pcDNA3.1-CD163 vector was constructed. The CD163 gene encoding 1115 amino acids was 3345 bp in length, and homologous comprison showed 99.37% with the sequence reptorted in GenBank (EU016226). The recombinant plasmid was transfected into PK-15 cells. At 72h hours after transfection, the expression of CD163 was confirmed by indirect immunofluorescence assay (IFA) with the monoclnal antibody of anti-CD163. The cells inoculated by PRRSV could be infected by IFA . |
| Key words: PAM PRRSV Scavenger Receptor CD163 |
