| TAT-HBX-EGFP蛋白的表达与纯化 |
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| 引用本文： | 刘宁,石英,魏征,陈德喜,娄金丽.TAT-HBX-EGFP蛋白的表达与纯化[J].胃肠病学和肝病学杂志,2011,20(1):41-43. |
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| 作者姓名： | 刘宁 石英 魏征 陈德喜 娄金丽 |
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| 作者单位： | 1. 首都医科大学附属北京佑安医院性病艾滋病实验室,北京,100069
2. 首都医科大学附属北京佑安医院性病艾滋病实验室,北京100069;北京市肝病研究所 |
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| 摘 要： | 目的构建TAT-HBx-EGFP质粒,转化大肠埃希菌BL21,进行蛋白的表达、鉴定及纯化。方法 PCR扩增目的基因HBx,进行EcolR1单酶切,与经EcolR1单酶切的TAT-EGFP质粒进行连接,转化大肠埃希菌BL21,利用IPTG对其进行诱导。应用West-ern blot方法鉴定表达的融合蛋白,采用Ni2＋-金属螯合亲和层析柱法对融合蛋白进行纯化。结果得到构建好的TAT-HBx-EGFP质粒及纯化的目的融合蛋白TAT-HBX-EGFP。结论通过对TAT-HBx-EGFP质粒的构建和TAT-HBX-EGFP蛋白的表达与纯化,为进一步研究HBX蛋白的致肝癌作用奠定了实验基础。
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| 关 键 词： | TAT-HBx-EGFP质粒 构建 融合蛋白 表达与纯化 |
The expression and purification of TAT-HBX-EGFP protein |
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| LIU Ning,SHI Ying,WEI Zheng,CHEN Dexi,LOU Jinli.The expression and purification of TAT-HBX-EGFP protein[J].Chinese Journal of Gastroenterology and Hepatology,2011,20(1):41-43. |
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| Authors: | LIU Ning SHI Ying WEI Zheng CHEN Dexi LOU Jinli |
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| Institution: | LIU Ning1,SHI Ying1,WEI Zheng1,CHEN Dexi1,LOU Jinli1,2 1.Laboratory of STD and AIDS,Beijing YouAn Hospital,Capital Medical University,Beijing 100069,2.Beijing Institute of Liver Diseases,China |
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| Abstract: | Objective To construct a TAT-HBx-EGFP plasmid transformed into E.coli BL21,then protein expression was identified and purified.Methods PCR was used to amplify gene HBx.HBx and TAT-EGFP plasmid were digested by EcolR1 single restriction enzyme,then joined together.The plasmid was transformed into E.coli BL21,and induced by IPTG.Western blot was used to identify the protein and purified by Ni2 +-metal chelate affinity chromatography.Results We got the constructed TAT-HBx-EGFP plasmid and the purified fusion T... |
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| Keywords: | TAT-HBx-EGFP plasmid Construction Fusion protein Expression and purification |
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