Marc-145细胞CD163基因克隆及其真核表达质粒构建
猪繁殖与呼吸综合征是危害养猪业重要传染病之一,CD163是猪繁殖与呼吸综合征感染易感细胞受体之一。从Marc-145细胞克隆出CD163基因并测序,再根据得到序列,应用Primer5.0软件设计引物,成功地扩增出带酶切位点CD163ORF基因,通过Hind11I和XhoI双酶切位点将CD163基因克隆入真核表达载体pcDNA3.1-myc—His中,获得重组质粒pcDNA3.1-CD163,并将重组质粒pcDNA3.1-CD163在BHK细胞上进行表达鉴定。结果表明,Marc-145细胞CD163蛋白在BHK细胞中得到了表达。这为进-步研究CD163受体在PRRSV感染Marc-145细胞过程...
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Published in | 东北农业大学学报 Vol. 44; no. 6; pp. 28 - 31 |
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Main Author | |
Format | Journal Article |
Language | Chinese |
Published |
华南农业大学农业部兽用疫苗创制重点实验室,广州510642%华南农业大学农业部兽用疫苗创制重点实验室,广州,510642%海南省农业科学院畜牧兽医研究所海南省热带动物繁育与疫病研究重点实验室,海口,571100
2013
海南省农业科学院畜牧兽医研究所海南省热带动物繁育与疫病研究重点实验室,海口571100 |
Subjects | |
Online Access | Get full text |
ISSN | 1005-9369 |
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Summary: | 猪繁殖与呼吸综合征是危害养猪业重要传染病之一,CD163是猪繁殖与呼吸综合征感染易感细胞受体之一。从Marc-145细胞克隆出CD163基因并测序,再根据得到序列,应用Primer5.0软件设计引物,成功地扩增出带酶切位点CD163ORF基因,通过Hind11I和XhoI双酶切位点将CD163基因克隆入真核表达载体pcDNA3.1-myc—His中,获得重组质粒pcDNA3.1-CD163,并将重组质粒pcDNA3.1-CD163在BHK细胞上进行表达鉴定。结果表明,Marc-145细胞CD163蛋白在BHK细胞中得到了表达。这为进-步研究CD163受体在PRRSV感染Marc-145细胞过程中作用以及与其他受体之间相互作用提供试验数据。 |
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Bibliography: | Porcine reproductive and respiratory syndrome virus (PRRSV) has been acknowledged as one of the most important agents affecting swine. The scavenger receptor CD163 is one of the important entry mediators for PRRSV. In order to ascertain the function of CD163 gene, the CD163 gene was cloned from Marc-145 cell by RT-PCR, and then the whole coding frame of CD163 was cloned into the eukaryotic expression vector pcDNA3.1-myc-His. Indirect fluorescence microscopy showed that CD163 were expressed in transfected cells. The results lay foundation for analyzing the function of CD163 in cultured cell. PRRSV; Marc-145 cell; CD163; cloning; expression vector 23-1391/S CAO Zongxi, JIAO Peirong, LIN Zhemin, YU Junyong, WU Xinwei, ZHANG Guihong(1. Hainan Provincial Key Laboratory of Tropical Animal Reproduction and Breeding and Veterinary Medicine, Institute of Animal Husbandry and Veterinary Medicine, Hainan Academy of Agricultural Sciences, Haikou 571100, China; 2. MOA Key laboratory for Animal Vaccine Development, South Chin |
ISSN: | 1005-9369 |