| 人单核细胞趋化蛋白1基因的克隆及在大肠杆菌中的表达 |
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| 引用本文： | 王汉涛,郭葆玉,屠岳. 人单核细胞趋化蛋白1基因的克隆及在大肠杆菌中的表达[J]. 第二军医大学学报, 2000, 21(4): 318-321 |
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| 作者姓名： | 王汉涛 郭葆玉 屠岳 |
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| 作者单位： | 第二军医大学长海医院普通外科!上海 200433,药学院生化药学教研室,第二军医大学长海医院普通外科!上海 200433,长征医院普通外科,基础医学部卫生毒理学教研室,第二军医大学长海医院普通外科!上海 200433 |
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| 基金项目： | 国家自然科学基金资助项目!（39770689） |
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| 摘 要： | 目的：克隆表达具有重要生物功能的人单核细胞核细胞趋化蛋白１（ｈｕＭＣＰ－１）。方法：从人外周血单核细胞（ＰＢＭＣ）中提取ｐｏｌｙ（Ａ）＾＋ＲＮＡ,用ＲＴ－ＰＣＲ法扩增出ｈｕＭＣＰ－１ｃＤＮＡ。将ｈｕＭＣＰ－１ｃＤＮＡ插入融合表达载体ｐＧＥＸＩＮ,１ｍｍｏｌ／Ｌ异丙基－β－Ｄ－硫代半乳糖苷（ＩＰＴＧ）诱导后获得高效表达。结果：Ｗｅｓｔｅｒｎ ｂｌｏｔ证明与ｈｕＭＣＰ－１单克隆抗体有明显交叉反应。结论
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| 关 键 词： | 基因克隆 融合表达 单核细胞趋化蛋白 huMCP－1 |
| 修稿时间： | 】 |
Cloning of human monocyte chemoattractant protein-1 and fusion expression in E. coli |
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| WANG Han-Tao, GUO Bac-Yu, TU Yue, WANG Yuan-He, ZHANG Shu-Ying, MENG Rong-Gui. Cloning of human monocyte chemoattractant protein-1 and fusion expression in E. coli[J]. Former Academic Journal of Second Military Medical University, 2000, 21(4): 318-321 |
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| Authors: | WANG Han-Tao GUO Bac-Yu TU Yue WANG Yuan-He ZHANG Shu-Ying MENG Rong-Gui |
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| Abstract: | Objective: To clone and express human monocyte chemoattractant protein-1 (huMCP-1 ) with important biologic functions. Methods: Poly (A)~+ RNA was extracted from PBMC and huMCP-1 cDNA was obtained by RT-PCR. huMCP-1 cDNA was inserted into fusion expression vector pGEXIN site of BamH I,QD and EcoR I,and gained high expres- sion induced by 1 mmol/L IPTG. Results: Western blotting analysis showed that the product had apparent cross reaction with huMCP-1 monoclonal antibody. Conclusion: Success on cloning and expression of huMCP-1 provide useful material for the laboratory and clinic research of MCP-1. |
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| Keywords: | monocyte chemoattractant protein-1 gene cloning fusion expression |
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