Vascular endothelial growth factors 165 (VEGF 165 ) is the most potent and widely used pro-angiogenic factor. Here we determined optimal culture condition of recombinant human VEGF 165 (rhVEGF 165 ) in Escherichia coli (E. coli). rhVEGF 165 expression was the highest in 0.25% of l-arabinose induction concentration, at 20°C induction temperature, and for 5h induction time under the control of araBAD promoter using pBADHisA vector. In biological activity test, rhVEGF 165 significantly increased the proliferative activity of CPAE cells (p<0.001) and upregulated the expressions of endothelial cell growth-related genes, such as platelet endothelial cell adhesion molecule (PECAM-1), endothelial-specific receptor tyrosine kinase (TEK), kinase insert domain protein receptor (KDR), and tyrosine kinase with immunoglobulin-like and EGF-like domains 1 (TIE1) in calf pulmonary artery endothelial (CPAE) cells.