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[Cloning of VEGF receptor KDR and its expression in insect cells].
- Source :
-
Sheng wu gong cheng xue bao = Chinese journal of biotechnology [Sheng Wu Gong Cheng Xue Bao] 2001 Mar; Vol. 17 (2), pp. 140-4. - Publication Year :
- 2001
-
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.
- Subjects :
- Angiogenesis Inhibitors pharmacology
Animals
Chick Embryo
Cloning, Molecular
Endothelium, Vascular cytology
Endothelium, Vascular drug effects
Humans
Receptor Protein-Tyrosine Kinases pharmacology
Receptors, Vascular Endothelial Growth Factor
Spodoptera
Baculoviridae genetics
Receptor Protein-Tyrosine Kinases genetics
Receptors, Growth Factor genetics
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1000-3061
- Volume :
- 17
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 11411219