›› 2008, Vol. 1 ›› Issue (7): 40-42.

• 生物技术 • 上一篇    下一篇

鸡传染性法氏囊病病毒VP2蛋白GFP融合分子在COS7细胞内的表达

李姣1,2,张红1,3,张改平1,王爱萍2,易明林1,郅玉宝1,鲁琨1,张丽萍1   

  1. 1. 河南省动物免疫学重点实验室,郑州 450002; 2. 郑州大学,郑州 450001;3. 河南教育学院,郑州 450003
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-07-20 发布日期:2008-07-20

Expression of VP2 Protein of Infectious Bursal Disease Virus Fused with GFP in COS7 Cells

LI Jiao1,2, ZHANG Hong1,3, ZHANG Gaiping1, WANG Aiping2, YI Minglin1, ZHI Yubao1, LU Kun1, ZHANG Liping1   

  1. Henan Key Laboratory for Animal Immunology, Zhengzhou 450002, China; 2.Zhengzhou University, Zhengzhou 450001, China; 3.Henan Educational Institute, Zhengzhou 450003, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-07-20 Published:2008-07-20

摘要: 将鸡传染性法氏囊病病毒(IBDV)VP2蛋白基因插入增强型绿色荧光表达载体pEGFP-C1中,构建了真核表达载体pEGFP-VP2。PCR与酶切鉴定结果表明VP2基因成功插入到表达载体pEGFP-C1中。脂质体法转染COS7细胞后,荧光显微镜检测到了GFP-VP2融合蛋白的表达。用200 μg/ml的G418成功筛选到了稳定表达GFP-VP2融合蛋白的细胞系,表达蛋白经镍亲和柱得到了纯化。

关键词: 鸡; 传染性法氏囊病病毒; VP2蛋白; 真核表达; GFP融合蛋白

Abstract: VP2 gene of infectious bursal disease virus(IBDV) was inserted into enhanced GFP expression vector pEGFP-C1. The recombinant eukaryotic expression vector pEGFP-VP2 was constructed, and detected by endonuclease digestion and PCR , and then was ransfected to COS7 cells by lipid reagent. After infecting, green fluorescence was observed under fluorescent microscope. The results indicates that GFP-VP2 fusion protein was successfully expressed in COS7 cells. Stable expressing cells was selected by 200 μg/ml G418, and GFP-VP2 fusion protain was purified by NiHis affinity column.

Key words: chicken; IBDV; VP2 protein; eukaryotic expression; GFP fusion protein

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