[Cloning of VEGF receptor KDR and its expression in insect cells].
The cDNA fragment of the first 3 loops of VEGF receptor, KDR, was cloned by PCR and inserted into a baculovirus expression plasmid pFASTBACI. The competent E. coli DH10BAC cell, which contain another plasmid with baculovirus genome in it, was transformed with pFASTBACI-KDRn3. Homologous recombination in the prokaryotic cells resulted in a recombinant plasmid containing KDRn3 in baculovirus genome. Transfection of the insect cell SF-9 with above plasmid generated a recombinant baculorvirus contain target gene fragment. SDS-PAGE and Western blot analysis of the supernatant of the infected SF-9 cell showed that KDRn3 was secreted in the medium. The recombinant protein was verified with Western blot and tested for their binding activity with VEGF. Its anti-angiogenic activity was assayed on chorionic allantoic membrane(CAM) of fertilized egg. The results showed that the recombinant protein could inhibit new vessel formation on CAM of fertilized eggs.
Zeng
G F
GF
State Key Laboratory of Molecular Virology and Genetic Engineering, Beijing 100052, China.
Zhang
Z Q
ZQ
Zhang
L G
LG
Chen
A J
AJ
Yao
L H
LH
Hou
Y D
YD
D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002642:Chick Embryo; D003001:Cloning, Molecular; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D006801:Humans; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics* / Q000494:pharmacology; D017978:Receptors, Growth Factor / Q000235:genetics*; D040262:Receptors, Vascular Endothelial Growth Factor; D018411:Spodoptera
D020533:Angiogenesis Inhibitors; D017978:Receptors, Growth Factor; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor