摘要
目的：克隆表达具有重要生物功能的人单核细胞趋化蛋白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：ｈｕＭＣＰ－１基因的克隆并表达成功为今后的实验室和临床研究提供了有用的材料。
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.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2000年第4期318-321,共4页
Academic Journal of Second Military Medical University
基金
国家自然科学基金资助项目!(39770689)
