Cloning of VEGF Receptor KDR and its Expression in Insect Cells
VEGF受体KDR胞外区基因的克隆及其在昆虫细胞中的表达

The cDNA fragment of the first 3 loops of VEGF receptor, KDR, was cloned by PCR and inserted into a baculovirus expression plasmid pF AST B AC I. The competent \%E.coli\% DH10BAC cell, which contain another plasmid with baculovirus genome in it, was transformed with pF AST B AC I\|KDR\-\{n3\}. Homologous recombination in the prokaryotic cells resulted in a recombinant plasmid containing KDR\-\{n3\} in baculovirus genome. Transfection of the insect cell SF\|9 with above plasmid genatated a recombinant baculorvirus contain target gene fragment. SDS\|PAGE and Western blot analysis of the supernatant of the infected SF\|9 cell showed that KDR\-\{n3\} 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.