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Cloning of human monocyte chemoattractant protein-1 and fusion expression in E. coli

WANG Han-Tao; GUO Bac-Yu; TU Yue; WANG Yuan-He; ZHANG Shu-Ying; MENG Rong-Gui (1. Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China; 2. Department of Bio- chemical Medicine, College of Pharmacy; 3  
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.
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