| 人组蛋白H1基因的原核表达、纯化及其活性检测 |
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| 作者姓名： | Xu XJ Fan ZY Wang LX Zhang H Ding LH DU N Ye QN |
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| 作者单位： | 1. 军事医学科学院生物工程研究所,北京,100850
2. 解放军总医院第一附属医院,北京,100048 |
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| 基金项目： | 全军医药卫生科研基金资助,国家重大科学研究计划,国家自然科学基金资助项目 |
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| 摘 要： | 目的:克隆人组蛋白H1全长编码区基因,获得其原核表达产物,并对融合蛋白进行纯化以及活性检测.方法:采用PCR技术从人乳腺文库中扩增出人组蛋白H1全长编码区基因,将其克隆到pGEX-KG载体中,在大肠杆菌Rossate中表达后,利用GST-Sepharose 4B亲和珠对原核表达产物进行纯化,以SDS-PAGE和Western blot鉴定表达与纯化产物.结果:从人乳腺文库中扩增获得约650 bp的DNA片段,并成功克隆至pGEX-KG载体上,经测序与目的序列完全一致;在Rossate菌中诱导表达出相对分子质量(Mr)约为52 000的目的蛋白,经纯化后获得了纯度较高的重组蛋白GST-H1,激酶实验证明该蛋白活性良好.结论:成功获得了活性良好的重组蛋白GST-H1,为后续研究细胞周期蛋白调控奠定了实验基础.
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| 关 键 词： | 人组蛋白H1 原核表达 纯化 |
Prokaryotic expression and purification of human histone-1 and its activity detection |
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| Xu XJ,Fan ZY,Wang LX,Zhang H,Ding LH,DU N,Ye QN.Prokaryotic expression and purification of human histone-1 and its activity detection[J].Journal of Cellular and Molecular Immunology,2011,27(8):843-845. |
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| Authors: | Xu Xiao-jie Fan Zhong-yi Wang Ling-xue Zhang Hao Ding Li-hua DU Nan Ye Qi-nong |
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| Institution: | Beijing Institute of Biotechnology, Beijing 100850, China. |
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| Abstract: | AIM: To construct the prokaryotic expression vector of human histone-1,obtain the purified GST-H1 protein,and detect its activity.METHODS: Human histone-1 coding region was amplified from human mammary cDNA library,and was inserted into prokaryotic expression vector pGEX-KG.The recombinant plasmid pGEX-KG-H1 was transformed into E.coli Rossate.The expressed product was purified by GST-Sepharose 4B beads and identified by SDS-PAGE and Western blot analysis.RESULTS: The DNA fragment of about 650 bp was succes... |
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| Keywords: | human histone-1 prokaryotic expression purification |
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