Objective:To make recombinant vascular endothelial growth factor(VEGF) in E.Coli. Methods:The VEGF cDNA was cloned and put into the bacterial expression system PET32a(Novagen) in frame with histidine tag. After introducing into the BL21 (DE3) E coli host,recombinant VEGF165 protein was produced and purified by denaturing condition on the His-bind resin. Results:The recombinant protein was confirmed by AA sequencing. The reconstituted VEGF165 was recognized by its receptor Neuropilin-1 and could obstruct the expression of Neuropilin-1 in cell curface. Conclusion:pET32a/human VEGF165 expression vector was transfected into E.coli BL21(DE3) and produced a lot of functional recombinant human VEGF165.