| TNFR1BP/IgGFc融合蛋白载体的构建、表达及其生物学活性研究 |
| |
| 引用本文： | 黄丽霞,尹丙姣,王晶,梁慧芳,曾庆岭,姜小丹,李卓娅.TNFR1BP/IgGFc融合蛋白载体的构建、表达及其生物学活性研究[J].华中科技大学学报(医学版),2008,37(5). |
| |
| 作者姓名： | 黄丽霞 尹丙姣 王晶 梁慧芳 曾庆岭 姜小丹 李卓娅 |
| |
| 作者单位： | 华中科技大学同济医学院基础医学院免疫学系,武汉,430030 |
| |
| 基金项目： | 国家自然科学基金，国家高技术研究发展计划(863计划) |
| |
| 摘 要： | 目的 构建肿瘤坏死因子受体1(TNFR1)封闭肽与IgGFc融合蛋白的真核表达载体,检测其表达和生物学活性.方法 将合成的TNFR1封闭肽(TNFR1BP)基因片段插入质粒pIG/3C,构建真核表达载体pIG/3C-TNFR1BP;脂质体法将重组载体转染COS-7细胞;ELISA和Western blot法检测转染细胞的培养上清中TNFR1BP/IgGFc融合蛋白的表达;间接免疫荧光抑制实验、细胞毒抑制实验检测融合蛋白对TNFR1的封闭作用.结果 经PCR和核苷酸序列测定证实TNFR1BP基因正确插入质粒pIG/3C.ELISA检测证实,在转染细胞培养上清中有融合蛋白的表达;间接免疫荧光抑制实验和细胞毒抑制实验证实融合蛋白能结合L929细胞表面的TNFR1,并能抑制分泌型肿瘤坏死因子α(sTNF-α)介导的细胞毒作用.结论 成功构建并实现TNFR1BP/IgGFc融合蛋白真核表达;体外实验证实此融合蛋白可通过与细胞表面TNFR1结合,从而拮抗sTNF-α介导的生物学效应.
|
| 关 键 词： | 肿瘤坏死因子-α 真核表达 IgGFc融合蛋白 封闭肽 |
Cloning and Expression of Human TNFR1BP/IgGFc Fusion Protein in COS-7 Cells |
| |
| Huang Lixia,Yin Bingjiao,Wang Jing et al.Cloning and Expression of Human TNFR1BP/IgGFc Fusion Protein in COS-7 Cells[J].Journal of Huazhong University of Science and Technology(Health Sciences),2008,37(5). |
| |
| Authors: | Huang Lixia Yin Bingjiao Wang Jing |
| |
| Institution: | Huang Lixia,Yin Bingjiao,Wang Jing et al Department of Immunology,School of Basic Medical Sciences,Tongji Medical College,Huazhong University of Science , Technology,Wuhan 430030 |
| |
| Abstract: | Objective To express the fusion protein of the TNFR1 blocking peptide with Fc fragment of human IgG1 in mammalian cells and identify the biological effect of its expressive product.Methods The eukaryotic cell expression vector pIG/3C-TNFR1BP was constructed by inserting the TNFR1 blocking peptide(TNFR1BP) gene into plasmid pIG/3C and transfected into COS-7 cells with Lipofectamine reagent.The expression of fusion protein(TNFR1BP/IgGFc) was tested by ELISA and Western blot.Inhibition of the fusion protein to... |
| |
| Keywords: | |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
