Objective To be expressed human vascular endothelial growth factor (VEGF) recombinant protein in Escherichia Coli in high level.
Methods VEGF was amplified from human fetal brain cDNA library, the amplified fragment was inserted into M13mP18 and confirmed to be VEGF165cDNA by restriction mapping and DNA sequencing, then it was combined with an expression vector PRL621. This recombinant plasmid overexpressed a 20kd recombinant protein in E.Coli(TG1), the protein was isolated and purifed from E.Coli, and initially renatured.
Results The overexpressed recombinant protein was 35% of the total cell protein, the sequence of its first 15-N terminal amino acid was identrical to that of the human natural VEGF protein, Chorioallantoic membrane(CAM) assay showed that the rhVEGF promated new capillary vessels formation.
Conclusion The genetic engineering Escherichia Coli can express human vascular endothelial growth factor in high level.
Citation： HE Yanzheng,LIU Jianping,XIE Yi,et al.. CLONING cDNA OF HUMAN VASCULAR ENDOTHELIAL GROWTH FACTOR AND HIGH LEVEL EXPRESSION IN ESCHERICHIA COLI. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2001, 8(2): 72-74. doi: Copy