| 带有穿膜肽重组PTD-Sox2的制备及活性鉴定 |
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| 引用本文： | 刘晓飞,余榕捷,王静静,苏航,黄霖,陈建苏.带有穿膜肽重组PTD-Sox2的制备及活性鉴定[J].生物技术,2011,21(3). |
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| 作者姓名： | 刘晓飞 余榕捷 王静静 苏航 黄霖 陈建苏 |
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| 作者单位： | 1. 暨南大学生物工程研究所,广东广州,510632
2. 暨南大学医学院眼科,广东广州,510632 |
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| 基金项目： | 国家自然科学基金项目资助Supported bv the National Natural Science Foundation of China |
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| 摘 要： | 目的:制备带有穿膜肽的重组蛋白PTD-Sox2并对其活性进行鉴定.方法:通过将Sox2基因与穿膜肽基因PTD融合,利用原核表达载体pKYB进行表达,用Ni柱制备、纯化融合蛋白;用Western blot对其进行鉴定;用FITC标记PTD-Sox2,与CHO细胞孵育,检测其穿膜能力;用FRET法检测转录因子Sox2结合目的DNA的活性.结果:构建了带有穿膜肽重组PTD-Sox2的原核表达菌,制备并纯化PTD-Sox2,对其活性鉴定.结论:PTD-Sox2能够穿过细胞膜与核膜,进入细胞核内,穿膜效率为40.86%,并且可与目的DNA进行特异结合,为蛋白体外诱导iPS的产生奠定基础.
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| 关 键 词： | 核转录因子 原核表达 |
Preparation and Activity Identification of Recombinant PTD-Sox2 |
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| LIU Xiao-fei,YU Rong-jie,WANG Jing-jing,SU Hang,HUANG Lin,CHEN Jian-su.Preparation and Activity Identification of Recombinant PTD-Sox2[J].Biotechnology,2011,21(3). |
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| Authors: | LIU Xiao-fei YU Rong-jie WANG Jing-jing SU Hang HUANG Lin CHEN Jian-su |
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| Institution: | LIU Xiao-fei1,YU Rong-jie1,WANG Jing-jing1,SU Hang2,HUANG Lin1,CHEN Jian-su2*(1.Bioengineering Institute of Jinan University,Guangzhou 510632,China,2.Ophthalmology Department of Medical College,Jinan University,China) |
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| Abstract: | Objective: To prepare recombinant protein PTD-Sox2 with cell penetrating peptide and identify its activity.Method:The Sox2 gene was expressed as a fusion protein PTD-Sox2 with PTD using prokaryotic expression vector pKYB.The fusion protein was purified by Ni affinity chromatography and identified by Western blot analysis.The penetrating ability of the fusion protein PTD-Sox2 into CHO cells was investigated using fluorescein(FITC) labeling method.And the bioactivity of PTD-Sox2 to bind to the target DNA sequ... |
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| Keywords: | PTD-Sox2 iPS FITC FRET |
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