| 人致肺纤维化因子的重组表达和抗体制备 | |
| 陈晓华 ; 蔡国平 ; CHEN Xiao-hua ; CAI Guo-ping | |
| 2010-06-10 ; 2010-06-10 | |
| 关键词 | 肺纤维化 几丁质酶 重组表达 多克隆抗体 lung fibrosis chitinase recombinant expression polyclonal antibodies R392.11 |
| 其他题名 | Recombinant expression of human fibrogenic factors and preparation of polyclonal antibodies |
| 中文摘要 | 目的:进行人致肺纤维化因子的原核和真核表达,并利用原核表达蛋白制备兔抗人致肺纤维化因子多克隆抗体。方法:人致肺纤维化因子为同一基因的一组可变剪切产物,将其共有cDNA序列(ChS)插入质粒pQE-30,构建ChS-pQE表达质粒,转化大肠杆菌M15,表达菌株经IPTG诱导,获得大量不可溶的包涵体蛋白。以之为抗原免疫新西兰大白兔,制备出多克隆抗体。对该多克隆抗体初步检测证明获得较高滴度和较高特异性的抗体,并用Western blot鉴定这组cDNA连接到pcDNA3.1D载体后在真核细胞的重组表达。结果:原核表达的不可溶性重组蛋白也能成功地制备出多克隆抗体。结论:成功地表达了人致肺纤维化因子,并制备出高滴度、高特异性的多克隆抗体,人致肺纤维化因子的表达和功能研究提供了一条途径。; AIM: To express the recombinant proteins of human fibrogenic factors and prepare the polyclonal antibodies of rabbit-anti-human fibrogenic factors. METHODS: The human fibrogenic factors were variant splicing forms of one gene. Their consensus sequence (ChS) was cloned into plasmid pQE30 to construct recombinant prokaryotic expression system. The recombinant expression vectors were transformed into E.coli M15, and then the expression vectors were induced by IPTG and many insoluble inclusion body proteins was obtained. The obtained proteins were used to immunize rabbits to obtain polyclonal antiserum with high titer and specificity. The eukaryotic expression of the recombinants of human fibrogenic factors was detected by Western blot. RESULTS: The insoluble inclusion body proteins were also used to prepare polyclonal antibodies with high titer and specificity. CONCLUSION: The expression of the recombinant proteins of human fibrogenic factors and the preparation of polyclonal antiserum provide a useful tool for further study of the expression and function of human fibrogenic factors.; 国家自然科学基金资助项目(30270515) |
| 语种 | 中文 ; 中文 |
| 内容类型 | 期刊论文 |
| 源URL | [http://hdl.handle.net/123456789/60615]  |
| 专题 | 清华大学 |
| 推荐引用方式 GB/T 7714 | 陈晓华,蔡国平,CHEN Xiao-hua,等. 人致肺纤维化因子的重组表达和抗体制备[J],2010, 2010. |
| APA | 陈晓华,蔡国平,CHEN Xiao-hua,&CAI Guo-ping.(2010).人致肺纤维化因子的重组表达和抗体制备.. |
| MLA | 陈晓华,et al."人致肺纤维化因子的重组表达和抗体制备".(2010). |
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