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| 肿瘤坏死因子相关凋亡诱导配体的表达及其活性检测 | |
| 引用本文： | 赵雪娜,武国栋,孙丽霞,黄山英,王庆民,薛彤彤,王晶翼. 肿瘤坏死因子相关凋亡诱导配体的表达及其活性检测[J]. 生物技术, 2011, 21(4). DOI: 10.3969/j.issn.1004-311X.2011.04.090 |
| 作者姓名： | 赵雪娜 武国栋 孙丽霞 黄山英 王庆民 薛彤彤 王晶翼 |
| 作者单位： | 齐鲁制药有限公司药物研究院,山东济南,250100 |
| 摘 要： | 目的:构建肿瘤坏死因子相关凋亡诱导配体(TRAIL)高表达工程菌株,以获得具有生物学活性的重组TRAIL蛋白.方法:合成TRAIL基因,转到大肠杆菌中,IPTG诱导获得成功表达；进一步优化,增加可溶形式的蛋白质.利用硫酸铵对发酵液进行沉淀,经Ni柱、阴离子和阳离子交换柱层析,最终获得蛋白质纯品,对其进行体外活性检测.结果:经过IPTG诱导后,TRAIL达到菌体总蛋白的30％；经过优化,可溶形式蛋白达到55％.纯化获得纯度高于95％的TRAIL样品,体外测活结果表明:TRAIL蛋白能有效抑制多种肿瘤细胞增殖,抑制率达到70％-92％.结论:重组TRAIL以可溶形式得到高效表达,且具有较好的生物学活性,为其开发成药用蛋白奠定基础. |
| 关 键 词： | TRAIL 可溶形式表达 活性检测 抑制率 |
Expression and Activity Assay of Tumor Necrosis Factor-related Apoptosis-inducing Ligand |
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| ZHAO Xue-na,WU Guo-dong,SUN Li-xia,HUANG Shan-ying,WANG Qing-min,XUE Tong-tong,WANG Jing-yi. Expression and Activity Assay of Tumor Necrosis Factor-related Apoptosis-inducing Ligand[J]. Biotechnology, 2011, 21(4). DOI: 10.3969/j.issn.1004-311X.2011.04.090 | |
| Authors: | ZHAO Xue-na WU Guo-dong SUN Li-xia HUANG Shan-ying WANG Qing-min XUE Tong-tong WANG Jing-yi |
| Affiliation: | ZHAO Xue-na,WU Guo-dong,SUN Li-xia,HUANG Shan-ying,WANG Qing-min,XUE Tong-tong,WANG Jing-yi*(Department of Research and Development,Qilu Pharmaceutical Co.Ltd.,Jinan 250100,China) |
| Abstract: | Objective: Construction of TRAIL expression of engineering strain in order to obtain biologically active recombinant protein.Method:TRAIL gene was expressed in E.coli successfully after IPTG induced.Then the soluble forms of protein was increased by Optimization of the host and the temperature.TRAIL was purified by ammonium sulphate precipitation,Ni Sepharose 4B,Q sepharose FF and Source 30 S and analyzed its activity in vitro.Result: TRAIL was up to 30% of the total bacterial protein after IPTG induction,f... |
| Keywords: | TRAIL soluble expression activity assay inhibition rate |
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