Objective: To construct the eukaryotic expression vector of monocyte chemokine protein 1 (MCP-1) driven by the cytomegalovirus (CMV) promoter, transfect the vector into HEK293T cells, and detect its chemotaxis to macrophages.
Methods: MCP-1 promoter was obtained from mouse genome by PCR, and inserted into the vector named pFLAG-CMV1. We validated it by double-enzymes digestion and sequencing. Then HEK293T cells were transfected with pFLAG-CMV1-MCP-1. The expression of MCP-1 was detected by Western blotting. Finally, we identified the chemotaxis of the recombinant vector to macrophage by Transwell(TM) assay.
Results: The recombinant vector could generate target fragment by double enzyme digestion. HEK293T cells expressed MCP-1 after they were transfected with the recombinant vector, which increased the migration of macrophages.
Conclusion: MCP-1 expressed by HEK293T cells could apparently increase the migration of macrophages.