| hLMO4基因重组质粒的构建及蛋白表达和定位 |
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| 引用本文： | 李彦姝,王春玉,苏楠,张红艳,李丹妮,李丰.hLMO4基因重组质粒的构建及蛋白表达和定位[J].解剖科学进展,2009,15(1). |
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| 作者姓名： | 李彦姝 王春玉 苏楠 张红艳 李丹妮 李丰 |
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| 作者单位： | 1. 中国医科大学细胞生物教研室,细胞生物学卫生部重点实验室,辽宁,沈阳,110001
2. 中国医科大学94期临床医学,辽宁,沈阳,110001 |
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| 基金项目： | 国家自然科学基金，教育部博士点基金 |
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| 摘 要： | 目的构建hLMO4真核表达载体并证实融合蛋白在细胞内表达及定位。方法提取人乳腺癌MCF-7细胞的mRNA,反转录为cDNA。PCR扩增hLMO4全长编码基因,亚克隆至pCDNA3.1-myc-hisA表达载体中。将构建的重组质粒测序并转染到胃癌细胞BGC823中,提取细胞蛋白进行Western blot检测。利用共聚焦激光扫描显微镜观察pEGFP-LMO4在乳腺癌MCF-7细胞内定位。结果hLMO4全长基因序列克隆到了真核表达载体pCDNA3.1-myc-hisA中,酶切鉴定片段为500bp。Western blot检测到了融合蛋白表达,分子量约为23KD。pEGFP-LMO4在乳腺癌MCF-7细胞内定位以细胞核为主,在细胞浆内少量表达。结论成功的构建了hLMO4全长基因真核表达载体,pEGFP-LMO4蛋白定位于乳腺癌细胞质和核内。
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| 关 键 词： | hLMO4 乳腺癌 Western blot |
Construction of Fusion Plasmid of Human LMO4 Gene and Its Recombinant Protein Expression and localization |
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| LI Yan-Shu,WANG Chun-Yu,SU Nan,ZHANG Hong-Yan,LI Dan-ni,LI Feng.Construction of Fusion Plasmid of Human LMO4 Gene and Its Recombinant Protein Expression and localization[J].Progress of Anatomical Sciences,2009,15(1). |
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| Authors: | LI Yan-Shu WANG Chun-Yu SU Nan ZHANG Hong-Yan LI Dan-ni LI Feng |
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| Institution: | 1.Department of Cell Biology;Key Laboratory of Cell Biology of Ministry of Public Health of China;2.94k;Faculty of Clinical Medicine;China Medical University;Liaoning Shenyang 110001 China |
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| Abstract: | Objective To construct the expression plasmid of human LIM domain Only 4(hLMO4) gene and identify its recombinant protein expression and localization.Methods Total RNA was extracted from human breast cancer MCF-7 cells.The hLMO4 coding sequence was amplified by polymerase chain reaction(PCR) method and subcloned into pCDNA3.1-myc-his A vector.After the target region was sequenced,the plasmid was transfected into BGC 823 cell lines.The expression of the recombinant plasmid in BGC 823 cells was proved by West... |
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| Keywords: | hLMO4 Westem blot |
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