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利用SUMO和Strep标签的两步法制备和纯化TRAIL蛋白的评价
发表时间：2016-05-15 浏览量：2554 下载量：929
| 全部作者： | 韩晓建，黄晶晶，郝智超，金艾顺 |
| 作者单位： | 哈尔滨医科大学基础医学院 |
| 摘 要： | 为获得高纯度和生物活性的TRAIL蛋白，探讨两步纯化方法制备肿瘤坏死因子相关凋亡诱导配体（tumor necrosis factor related apoptosis-inducing ligand，TRAIL），并对纯度和活性进行评价。首先构建pSUMO-Strep-hTRAIL质粒，转化到原核细胞获得表达菌液，然后通过Ni+柱和Strep两步亲和层析的方法，简便、低成本地获得高产量和高纯度TRAIL蛋白。为评价TRAIL活性，检测TRAIL诱导细胞凋亡效果。结果显示，两步纯化法制备的TRAIL蛋白有较高纯度和生物学活性，可为体内抗肿瘤实验研究提供材料。 |
| 关 键 词： | 生物工程学；纯化方法；生物学活性；肿瘤坏死因子相关凋亡诱导配体 |
| Title： | Evaluation of SUMO-and Strep-tagged TRAIL protein prepared by two step of purification method |
| Author： | HAN Xiaojian, HUANG Jingjing, HAO Zhichao, JIN Aishun |
| Organization： | School of Basic Medical Sciences, Harbin Medical University |
| Abstract： | In order to obtain tumor necrosis factor related apoptosis-inducing ligand (TRAIL) protein with the high purity and biological activity, a two-step purification method was prepared using SUMO and Strep tagged, and the purity and activity of TRAIL were evaluated. Firstly, pSUMO-Strep-hTRAIL plasmid was constructed and transfected into prokaryotic cells to produce bacterial fluid, and then TRAIL protein of high purification was obtained by Ni+ and Strep using a two-step affinity chromatography. In order to evaluate the biological activity of TRAIL, the effect of TRAIL on inducing apoptosis was detected. The results showed that the TRAIL protein prepared by established method has good biological activity, which provides the material for anti-tumor experiments in vivo. |
| Key words： | biotechnology; purification method; biological activity; tumor necrosis factor related apoptosis-inducing ligand |
| 发表期数： | 2016年5月第9期 |
| 引用格式： | 韩晓建，黄晶晶，郝智超，等. 利用SUMO和Strep标签的两步法制备和纯化TRAIL蛋白的评价[J]. 中国科技论文在线精品论文，2016，9（9）：868-874. |
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