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目的 克隆获得编码人补体膜辅助调节蛋白（ ＭＣＰ） 的ｃＤＮＡ，并对其在真核细胞的表达及功能进行研究。方法 应用ＲＴＰＣＲ 方法，从Ｕ９３７ 细胞总ＲＮＡ 中扩增编码人ＭＣＰ 分子的ｃＤＮＡ片段， 快速克隆于ｐＧＥＭＴＥａｓｙ 载体，测定其序列。将该片段重组于ｐＬＸＳＮ 载体，电穿孔转染ＮＩＨ３Ｔ３ 细胞，经ＦＡＣＳ 检测筛选表达ＭＣＰ 的阳性细胞克隆，用补体溶破试验鉴定其抑制人补体溶破的功能。结果 ＲＴＰＣＲ 扩增得到１ １４４ｂｐ 的编码人ＭＣＰ 分子的ｃＤＮＡ 片段，序列分析表明该ｃＤＮＡ编码的蛋白为ＳＴＣ＋ ＣＹＴ２ 亚型。细胞转染筛选获得多个表达ＭＣＰ 的ＮＩＨ３Ｔ３ 阳性细胞克隆，补体溶破试验证实其具有抑制人补体经典途径和旁路途径溶破的功能。结论 本研究为进一步探讨不同亚型结构的ＭＣＰ 分子与功能的关系及其应用奠定了基础。
Objective To clone cDNA coding human complement membrane regulatory protein (MCP) and study its expression and function in eukaryotic cells. Methods RT-PCR method was used to amplify cDNA fragment encoding human MCP from total RNA of U937 cells and cloned into pGEM-Easyasy vector to determine the sequence. The fragment was recombined in pLXSN vector and transfected into NIH3T3 cells by electroporation. Positive cell clones expressing MCP were screened by FACS, and its complement inhibitory activity was used to identify the function of human complement complement. Results RT-PCR amplified 1 144bp cDNA encoding human MCP molecule. Sequence analysis showed that the protein encoded by this cDNA was STC + CYT2 subtype. A number of NIH3T3-positive cell clones expressing MCP were obtained by cell transfection and screening. Complement lysis assay confirmed that they have the function of inhibiting the classical complement pathway and the bypass pathway. Conclusion This study laid the foundation for further exploration of the relationship between MCP molecules and their functions and their applications in different subtype structures.