| 摘要: |
| 为获得猪繁殖与呼吸综合征病毒（PRRSV）GP5蛋白，本研究以一株PRRSV NADC30-like毒株为模板，利用RT-PCR扩增出ORF5基因后，将该基因克隆至原核表达载体pCold-TF中，构建了重组质粒pCold-TF-GP5，转化Rosetta （DE3）后用IPTG诱导，经SDS-PAGE、Western blot鉴定，重组蛋白获得了可溶性表达，大小约为72ku。将该蛋白过镍柱纯化后获得浓度为0.5mg/ml的纯化蛋白，为下一步GP5蛋白单克隆抗体的制备奠定物质基础。 |
| 关键词: 猪繁殖与呼吸综合征病毒 GP5蛋白 原核表达 |
| DOI： |
| 投稿时间：2020-12-13修订日期：2021-02-01 |
| 基金项目:中国兽医药品监察所所级课题（201802） |
|
| Prokaryotic Expression and Identification of Porcine Reproductive and Respiratory Syndrome Virus GP5 Protein |
|
| (China Institute of Veterinary Drug Control) |
| Abstract: |
| In order to obtain the GP5 protein of porcine reproductive and respiratory syndrome virus (PRRSV), the ORF5 gene was amplified by RT-PCR using the PRRSV NADC30-like strain genomic RNA as the template. The ORF5 gene was cloned into the prokaryotic expression vector pCold-TF. The recombinant plasmid pCold-TF-GP5 was constructed and transformed into Rosetta（DE3）and induced by IPTG. The SDS-PAGE and Western blot identification indicates the recombinant protein was soluble and its size was about 72 ku. The protein was purified by nickel column with a concentration of 0.5mg/mL, which laid the material foundation for the preparation of GP5 monoclonal antibody in the next step. |
| Key words: PRRSV GP5 protein prokaryotic expression |
