Cloning, purification and biological activity of human vascular endothelial growth factor fragment in E. coli
Li, X.; Zeng, W.; Zhang, Y.
Zhonghua Zhong Liu Za Zhi 24(5): 448-450
2002
ISSN/ISBN: 0253-3766 PMID: 12485495 Document Number: 548660
Objective: To observe the effect of human vascular endothelial growth factor (VEGF) fragment (3apprx4 exon) in E. coli on anti-angiogenesis. Methods: Through RT-PCR amplification, endonuclease cut and DNA sequence analysis identification, hVEGF fragment cDNA was inserted into E. coli expression vector pTrcHis2A. The prokaryotic expression plasmid pTrcHis2A/VEGF3apprx4 was constructed and transformed into TOP1OF. Results: After 8hr isopropy-beta-D-thiogalactoside (IPTG) induction, VEGF fragment was expressed in 15% of total proteins through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The expressed protein was highly antigenic and specific. The VEGF fragment was further purified by affinity, which could inhibit HUVEC proliferation and neovascularization of the chick chorioallantoic membrane. Conclusion: VEGF fragment is anti-angiogenetic, which may potentially be used in oncologico-biological targeting therapy.