title: [Cloning of VEGF receptor KDR and its expression in insect cells].
abstract: 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.
authors: State Key Laboratory of Molecular Virology and Genetic Engineering, Beijing 100052, China.
authors: G F
authors: GF
authors: Zeng
authors: Z Q
authors: ZQ
authors: Zhang
authors: L G
authors: LG
authors: Zhang
authors: A J
authors: AJ
authors: Chen
authors: L H
authors: LH
authors: Yao
authors: Y D
authors: YD
authors: Hou
mesh_terms: 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
chemical_list: D020533:Angiogenesis Inhibitors; D017978:Receptors, Growth Factor; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor