PRRSV清道夫受体CD163基因的真核表达和功能鉴定  

The Eukaryotic Expression and Functional Identification of Scavenger Receptor CD163 of PRRSV

作　　者：李红[1] 周恩民[2] 刘成倩[1] 易建中[1] 

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106 [2]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《中国兽药杂志》2013年第10期10-14,共5页Chinese Journal of Veterinary Drug

基　　金：上海市科技兴农重点攻关项目(沪农科攻字(2007)第12-1号)

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。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 .

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 

分 类 号：S852.4[农业科学—基础兽医学]