【摘要】 人膜辅基蛋白MCP(membranecofactorprotein)和降解加速因子DAF(decay acceleratingfactor)是免疫排斥反应中补体系统活性调节的重要蛋白.以PCR的方法分别从成人肝cDNA文库和胎儿骨髓cDNA文库中扩增得到MCP和DAF基因,克隆入载体pUC19,再以所构建质粒为模板,以PCR的方式在二基因间加入编码柔性连接肽的DNA序列,构建融合基因MCP DAF,并克隆入pUC19质粒.将MCP、DAF及MCP DAF亚克隆至大肠杆菌表达载体pET 32a(+)或pET 32b(+),构建表达载体MCP pET32a,DAF pET32b及MCP DAF pET32a,分别导入E.coliBL21菌株中表达,得到预期分子质量大小的蛋白.更多还原

【Abstract】 Human membrane cofactor protein (MCP) and decayaccelerating factor (DAF) play important roles in human complement system activity regulation during rejection. MCP and DAF gene were amplified from adult liver cDNA library and infant marrow cDNA library using PCR and cloned into pUC19 plasmid. A fusion gene was constructed using these plasmid as templates through PCR reaction and cloned into pUC19 plasmid. The three genes were subcloned into expression vector pET32a(+) or pET32b(+) and expressed in Escherichia. coli.更多还原