[Cloning, purification and biological activity of human vascular endothelial growth factor fragment in E. coli].
OBJECTIVE
To observe the effect of human vascular endothelial growth factor (VEGF) fragment (3 approximately 4 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/VEGF(3 approximately 4) was constructed and transformed into TOP10F.


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.


CONCLUSIONS
VEGF fragment is anti-angiogenetic, which may potentially be used in oncologico-biological targeting therapy.
Li
Xianmao
X
Instititue of Gastroenterology, Nanfang Hospital, First Military Medical University of PLA, Guangzhou 510515, China.
Zeng
Weisen
W
Zhang
Yali
Y
D020533:Angiogenesis Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D003001:Cloning, Molecular; D016228:Endothelial Growth Factors / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008222:Lymphokines / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D010446:Peptide Fragments / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D020533:Angiogenesis Inhibitors; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D010446:Peptide Fragments; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D007544:Isopropyl Thiogalactoside