| PRRSV清道夫受体CD163基因的真核表达和功能鉴定 |
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| 引用本文： | 李 红,易建中.PRRSV清道夫受体CD163基因的真核表达和功能鉴定[J].中国兽药杂志,2013,47(10):10-14. |
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| 作者姓名： | 李 红 易建中 |
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| 作者单位： | 上海市农业科学院畜牧兽医研究所,上海市农业科学院畜牧兽医研究所 |
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| 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。
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| 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 |
| 收稿时间： | 2013/6/29 0:00:00 |
| 修稿时间： | 2013/8/20 0:00:00 |
The Eukaryotic Expression and Functional Identification of Scavenger Receptor CD163 of PRRSV |
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| Institution: | Animal Huabandry and Veterinary Research Institute,Shanghai Academy of Agricultural Science, |
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| 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 . |
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