| hLMO3基因全长及截短序列原核表达载体的构建及蛋白诱导表达 |
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| 引用本文： | 刘彤,顾卉,李洋,李丰.hLMO3基因全长及截短序列原核表达载体的构建及蛋白诱导表达[J].解剖科学进展,2012,18(1):8-11. |
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| 作者姓名： | 刘彤 顾卉 李洋 李丰 |
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| 作者单位： | 1. 中国医科大学 基础医学院细胞生物教研室教育部医学细胞生物学重点实验室
2. 中国医科大学 附属盛京医院中心实验室,沈阳,110001 |
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| 基金项目： | 国家自然科学基金重大研究计划 |
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| 摘 要： | 目的构建hLMO3原核全长及5个截短表达载体并鉴定融合蛋白的表达。方法提取工具细胞HEK293的mRNA,反转录为cDNA,PCR扩增hLMO3全长及截短编码基因,亚克隆至GST融合表达载体pGEX-5X-2中。在E.coli BL21中诱导GST-hLMO3全长及截短融合蛋白表达,并经免疫印迹鉴定结果。结果 hLMO3全长及5个截短基因序列克隆到了原核表达载体pGEX-5X-2中,并经测序鉴定正确。Westernblot在E.coli Bl21中检测到了相应融合蛋白的表达。结论成功构建hLMO3全长及5个截短基因序列原核表达载体,在E.coli BL21中诱导表达出GST-LMO3全长及截短融合蛋白。
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| 关 键 词： | hLMO3 工具细胞HEK293 原核表达 截短蛋白 |
Construction of prokaryotic plasmid of human LMO3 and deletions gene and identification of their recombinant proteins |
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| LIU Tong
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GU Hui
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LI Yang
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LI Feng.Construction of prokaryotic plasmid of human LMO3 and deletions gene and identification of their recombinant proteins[J].Progress of Anatomical Sciences,2012,18(1):8-11. |
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| Authors: | LIU Tong GU Hui LI Yang LI Feng |
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| Institution: | 1*(1.Research Center for Cell Biology,Key Laboratory of Medical Cell Biology Ministry of Public Education of China;2.Key labaoratory of Shengjing Hospital,China Medical University,Shenyang 110001 China) |
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| Abstract: | Objective To construct an prokaryotic expression vector of LMO3(LIM-domain only 3) full-length and deletion gene and identify their recombinant protein expressions induced by isopro-pyl-1-thio-b-Dgalactopyranoside(IPTG).Methods Total RNA was extracted from HEK293,hLMO3 and deletion coding sequences were amplified by polymerase chain reaction(PCR)method and cloned into pGEX-5X-2.The expression of GST-hLMO3 fusion proteins was induced by IPTG and identified by Western blot.Results The coding sequence of hLMO3 full-length and deletion gene were cloned into the pGEX-5X-2 which was transformed into BL21 successfuly,and identified by double enzymes digestion and sequencing.The expression of GST-hLMO3 fusion proteins induced by IPTG was observed in Bl21.Conclusion The recombinant prokaryotic plasmids were constructed successfully into pGEX-5X-2 and the expression of GST-LMO3 full-length and deletion fusion proteins was identified. |
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| Keywords: | LIM-domain only 3 HEK293 prokaryotic expression deletions |
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