| 人MCP cDNA在NIH3T3细胞中表达及功能研究 |
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| 引用本文： | 黄健, 李文鑫, 郑从义,. 人MCP cDNA在NIH3T3细胞中表达及功能研究[J]. 生物工程学报, 2000, 16(2): 155-157 |
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| 作者姓名： | 黄健 李文鑫 郑从义 |
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| 作者单位： | 武汉大学生命科学学院武汉 430072 |
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| 基金项目： | 湖北省"九五”重点科技计划项目资助课题(96192004). |
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| 摘 要： | 通过构建表达人膜辅因子蛋白ＭＣＰ编码区全长ｃＤＮＡ的重级载体ｐｃＤＮＡ３ＭＣＰ,以磷酸钙沉淀法转染ＮＩＨ３Ｔ３细胞,用Ｇ４１８筛选阳性克隆,并鉴定ＭＣＰ ｃＤＮＡ在细胞中的表达。将血清梯度稀释并与细胞孵育,然后检测细胞活力：灭活的人血清不能引起对照细胞与ＭＣＰ转染细胞的胞毒作用,而新鲜人血清可使对照细胞发生补体依赖的细胞毒反应,ＭＣＰ转染细胞由于ＭＣＰ蛋白的合成,细胞受到保护不发生该反应。另外,Ｍ
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| 关 键 词： | 同源限制因子 MCP NIG3T3细胞 补体溶破 表达 |
| 文章编号： | 1000-3061(2000)02-0155-03 |
| 修稿时间： | 1999-03-15 |
Studies on Expression and Function of Human Membrane Cofactor Protein in NIH3T3 Cells |
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| HUANG Jian LI Wen Xin ZHEN Cong Yi. Studies on Expression and Function of Human Membrane Cofactor Protein in NIH3T3 Cells[J]. Chinese journal of biotechnology, 2000, 16(2): 155-157 |
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| Authors: | HUANG Jian LI Wen Xin ZHEN Cong Yi |
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| Affiliation: | College of Life Sciences, Wuhan University. |
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| Abstract: | One recombinant human membrane cofactor protein (MCP) expression plasmid pcDNA3 MCP was constructed.After transfected into NIH3T3 cells with calcium phosphate DNA precipitate method,the expression clones were obtained by G418 selection.To determine whether MCP generated on heterologous cells by cDNA transfection retains its capacity to inhibit human complement,NIH3T3 cells transfected with MCP cDNA were incubated with human serum as well as guinea pig,rabbit serum.Under identical conditions,the control cells transfected with pcDNA3 vector were completely lysed by human serum but the cells transfected with MCP were barely lysed.However,the cells transfected with MCP were lysed by guinea pig and rabbit serum complement as the same extent as control cells. |
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| Keywords: | Expression of MCP cDNA NIH3T3 cell complement lysis |
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