在大肠杆菌中融合高效表达人MCP-1

High level expression of MCP-1 in the form of fusion protein in E.coli

目的：利用含强启动子的融合表达载体ｐＧＥＸ－２Ｔ对编码人单核细胞趋化蛋白－１（ＭＣＰ－１）的基因进行高效表达。方法：基因重组技术及蛋白纯化技术。结果：ＳＤＳ－ＰＡＧＥ显示表达产物占菌体蛋白的５０％ 。Ｗｅｓｔｅｒｎ Ｂｌｏｔ检测表明，表达产物可与抗ＭＣＰ－１ 抗体特异反应。生物学活性测定结果表明，表达产物具有明显的单核细胞趋化活性。结论：融合蛋白对ＭＣＰ－１

Objective:A new overexpression plasmid pGEX 2T/MCP 1 was constructed,in which a gene fragement encoding human monocyte chemoattractant protein 1 (MCP 1) was inserted downstream of tac promoter.Methods:DNA recombinant technique was used.Results:The expression level was 50% of total bacterial proteins by SDS PAGE.Western blot analysis showed that the expressed products reacted specifically with anti MCP 1 antibodies.Detection of activity by the method of agarose plates showed that the expression product had obvious monocyte chemoattractant activity.Conclusion:Fusion protein may not affect the chemoattractant activity of MCP 1.