| 人BTLA-Fc融合蛋白的制备及其生物学活性的研究 |
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| 引用本文： | 朱耿超,黄子逸,曹丽娟,陈永井,王雪峰,张学光. 人BTLA-Fc融合蛋白的制备及其生物学活性的研究[J]. 现代免疫学, 2011, 0(5) |
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| 作者姓名： | 朱耿超 黄子逸 曹丽娟 陈永井 王雪峰 张学光 |
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| 作者单位： | 苏州大学医学生物技术研究所;苏州大学基础医学与生物科学学院; |
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| 基金项目： | 国家自然科学基金资助项目(30600548); 苏州大学博士基金资助项目(Q4134801); 江苏省青蓝工程资助 |
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| 摘 要： | 为建立CHO/BTLA-Fc基因转染细胞株,使之能稳定分泌BTLA-Fc融合蛋白,采用PCR法从本单位构建的pEGZ-Term/B7-H1-Fc重组质粒中扩增人IgG1(Fc)恒定区,Xho I、BamH I双酶切后与真核表达载体pIRES2-EGFP连接为重组质粒pIRES2-EGFP-Fc,同时PCR法从pEGZ-Term/BTLA重组质粒中获得人BTLA胞外段基因片段,用Nhe I、Xho I双酶切插入上述pIRES2-EGFP-Fc构建重组载体pIRES2-EGFP/BTLA-Fc。脂质体法以该重组载体转染鼠CHO细胞,G418筛选并亚克隆化。Protein G亲和层析柱对上清中的融合蛋白进行纯化,Western blot作定性分析。CCK-8检测BTLA-Fc融合蛋白对抗人CD3单抗激发的T淋巴细胞增殖的影响。结果表明,基因转染CHO细胞培养上清中表达的BTLA-Fc融合蛋白能与L929/HVEM细胞有效结合,纯化的融合蛋白经Western blot鉴定有清晰的目的条带,而且BTLA-Fc融合蛋白对T细胞的体外增殖具有抑制作用。
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| 关 键 词： | B、T淋巴细胞衰减子(BTLA) 融合蛋白 单纯疱疹病毒侵入介质(HVEM) T细胞 |
Preparation of human BTLA-Fc fusion protein and study of its biological function |
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| ZHU Geng-chao,,HUANG Zi-yi,CAO Li-juan,CHEN Yong-jing,WANG Xue-feng,ZHANG Xue-guang. Preparation of human BTLA-Fc fusion protein and study of its biological function[J]. Current Immunology, 2011, 0(5) |
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| Authors: | ZHU Geng-chao HUANG Zi-yi CAO Li-juan CHEN Yong-jing WANG Xue-feng ZHANG Xue-guang |
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| Affiliation: | ZHU Geng-chao1,2,HUANG Zi-yi1,CAO Li-juan1,CHEN Yong-jing1,WANG Xue-feng1,ZHANG Xue-guang1,2(1.Institute for Medical Biotechnology,2.School of Biology and Basic Medical Sciences,Soochow University,Suzhou 215006,China) |
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| Abstract: | To establish the CHO/BTLA-Fc gene transfected cell line expressing human BTLA-Fc fusion protein encoding human IgG1(Fc) was amplified by PCR from recombinant vector pEGZ-Term/B7-H1-Fc and then the target fragment was inserted to eukaryotic expression vector pIRES2-EGFP after being digested with Xho I,BamH I.The extracellular grne of BTLA was amplified by PCR from recombinant vector pEGZ-Term/BTLA,inserted into recombinant vector pIRES2-EGFP-Fc after being digested with Nhe I,Xho I.The recombinant vector was... |
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| Keywords: | BTLA HVEM fusion protein T cell |
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