| pGEX-5X-1-hLMO4原核质粒构建及重组蛋白表达 |
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| 引用本文： | 张红艳,李彦姝,姚远,王春玉,王迪,李丰.pGEX-5X-1-hLMO4原核质粒构建及重组蛋白表达[J].东南大学学报(医学版),2012,31(2):139-143. |
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| 作者姓名： | 张红艳 李彦姝 姚远 王春玉 王迪 李丰 |
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| 作者单位： | 1. 中国医科大学,细胞生物学教研室,卫生部细胞生物学重点实验室,教育部医学细胞生物学重点实验室,辽宁,沈阳,110001
2. 辽宁省人民医院,消化内科,辽宁,沈阳,110016 |
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| 基金项目： | 国家自然科学基金资助项目,博士点基金资助项目 |
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| 摘 要： | 目的:构建hLMO4的原核表达载体并诱导、纯化和鉴定其表达。方法:hLMO4全长编码基因经BamHⅠ和XhoⅠ双酶切后,克隆至GST融合表达载体pGEX-5X-1,异丙基β-D硫代半乳糖苷(IPTG)在BL21大肠杆菌中诱导GST-hLMO4融合蛋白表达,利用Glutathione Sepharose 4B纯化诱导的融合蛋白,并经Western blot鉴定结果。结果:hLMO4编码序列克隆至pGEX-5X-1载体中,双酶切鉴定片段大小为500 bp,在E.coli BL21中IPTG诱导融合蛋白的表达,分子质量约为49 000 Da,成功纯化出GST及GST-hLMO4蛋白,Western blot检测到蛋白表达。结论:构建了hLMO4基因原核表达载体,鉴定了GST-hLMO4融合蛋白表达。
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| 关 键 词： | LMO4 蛋白纯化 融合蛋白 |
Construction of pGEX-5X-1-hLMO4 prokaryotic plasmidand identification of its recombinant protein |
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| ZHANG Hong-yan
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LI Yan-shu
,
YAO Yuan
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WANG Chun-yu
,
WANG Di
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LI Feng.Construction of pGEX-5X-1-hLMO4 prokaryotic plasmidand identification of its recombinant protein[J].Journal of Southeast Univ: Medical Sci Ed,2012,31(2):139-143. |
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| Authors: | ZHANG Hong-yan LI Yan-shu YAO Yuan WANG Chun-yu WANG Di LI Feng |
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| Institution: | 1(1.Department of Cell Biology,China Medical University;Key Laboratory of Cell Biology,Ministry of Public Health;Key Laboratory of Cell Biology,Ministry of Education,Shenyang 110001,China;2.Department of Gastroenterology,the People’s Hospital of Liaoning Province,Shenyang 110016,China) |
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| Abstract: | Objective:To construct prokaryotic expression vector of human LMO4 gene and induce,purify and identify its recombinant protein expression.Methods:The hLMO4 coding sequence was digested with BamHⅠ and XhoⅠ enzymes,and cloned into pGEX-5X-1.The expression of GST-hLMO4 fusion protein was induced by IPTG and identified by Western blot.Results:The coding sequence of hLMO4 gene was cloned into the pGEX-5X-1 plasmid which was transformed into E.coli BL21.The length of fragment was 500 bp.The expression of GST-hLMO4 fusion protein was induced by IPTG,and the molecular weight of protein was 49 000 Da.Conclusion:The recombinant prokaryotic plasmid was successfully constructed into pGEX-5X-1.The expression of GST-LMO4 fusion protein was induced by IPTG and identified. |
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| Keywords: | LMO 4 protein purification fusion protein |
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