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| 核心组蛋白的融合表达和纯化 | |
| 引用本文： | 曹佳,丁丽华,范忠义,程龙,蒋凯,徐晓洁,韩永健,徐承水,叶棋浓.核心组蛋白的融合表达和纯化[J].生物技术通讯,2011,22(4):484-487. |
| 作者姓名： | 曹佳 丁丽华 范忠义 程龙 蒋凯 徐晓洁 韩永健 徐承水 叶棋浓 |
| 作者单位： | 1. 军事医学科学院生物工程研究所,北京,100850;曲阜师范大学生命科学学院,山东,曲阜,273165 2. 军事医学科学院生物工程研究所,北京,100850 3. 曲阜师范大学生命科学学院,山东,曲阜,273165 |
| 摘 要： | 目的:克隆核心组蛋白H2A、H2B、H3和H4的基因,表达并纯化组蛋白与谷胱甘肽S-转移酶(GST)的融合蛋白。方法:用PCR方法从乳腺文库中扩增核心组蛋白H2A、H2B、H3和H4的编码序列,分别将其以正确相位与pGEX-KG载体中的GST编码序列融合,得到重组质粒pGST-H2A、pGST-H2B、pGST-H3和pGST-H4,分别转化大肠杆菌BL21,表达融合蛋白GST-H2A、GST-H2B、GST-H3和GST-H4;用谷胱甘肽-Sepharose 4B亲和纯化融合蛋白;用Western印迹检测融合蛋白的表达及纯化。结果:分别构建了核心组蛋白H2A、H2B、H3和H4的融合表达载体;Western印迹检测表明,融合蛋白GST-H2A、GST-H2B、GST-H3和GST-H4获得表达及纯化。结论:表达并纯化了H2A、H2B、H3和H4的融合蛋白,为进一步研究核心组蛋白的功能奠定了基础。 |
| 关 键 词： | 组蛋白 克隆 表达 融合蛋白 |
Fusion Expession and Purification of Nucleosmal Histones |
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| CA Jia,DING Li-Hua,FAN Zhong-Yi,CHENG Long,JIANG Kai,XU Xiao-Jie,HAN Yong-Jian,XU Cheng-Shui,YE Qi-Nong.Fusion Expession and Purification of Nucleosmal Histones[J].Letters in Biotechnology,2011,22(4):484-487. | |
| Authors: | CA Jia DING Li-Hua FAN Zhong-Yi CHENG Long JIANG Kai XU Xiao-Jie HAN Yong-Jian XU Cheng-Shui YE Qi-Nong |
| Institution: | CAO Jia1,2,DING Li-Hua1,FAN Zhong-Yi1,CHENG Long1,JIANG Kai1,XU Xiao-Jie1,HAN Yong-Jian1,XU Chneng-Shui2,YE Qi-Nong1 1.Beijing Institute of Biotechnology,Beijing 100850,2.Department of Biological Science,Normal University of Qufu,Qufu 273165,China |
| Abstract: | Objective: To clone,express and purify the nucleosmal histone family members H2A,H2B,H3 and H4.Methods: The coding sequences of H2A,H2B,H3 and H4 were amplified by PCR and fused in frame with the coding region of GST in the pGEX-KG vector to generate the pGST-H2A,pGST-H2B,pGST-H3 and pGST-H4 recombinant plasmids.The GST-H2A,GST-H2B,GST-H3 and GST-H4 fusion proteins were expressed in E.coli BL21 and purified by glutathione-Sepharose 4B affinity chromatography.The fusion proteins were characterized by Western... |
| Keywords: | nucleosmal histones cloning expression fusion protein |
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