Identification of polymorphisms within the vascular endothelial growth factor (VEGF) gene: Correlation with variation in VEGF protein production

Dysregulated vascular endothelial growth factor (VEGF) expression has been implicated as a major contributor to the development of a number of common disease pathologies. The aim of this study was to establish the extent of g enetic variability within the VEGF gene and to determine whether this genet ic variation influenced levels of VEGF protein expression. The promoter reg ion and exon 1 of the VEGF gene were screened for polymorphisms using singl e-stranded conformation (SSCP) polymorphism analysis and direct PCR-sequenc ing. We identified 15 novel sequence polymorphisms most of which were rare. Eleven of these polymorphisms were single base substitutions, three were s ingle base insertions and one was a two base deletion. Thirteen of the poly morphisms were located within the promoter and two in the 5' untranslated r egion (5'UTR) of the gene. We established PCR-RFLP typing systems for ten o f the polymorphisms. For the two common polymorphisms at - 460 and +405, we developed a combined sequence specific priming (SSP) PCR typing system to determine the cis/trans orientation of each allele and hence, ascertain hap lotypes, A significant correlation was observed between lipopolysaccharide (LPS) stimulated peripheral blood mononuclear cell (PBMC) VEGF protein prod uction and genotype for the +405 polymorphism. (C) 2000 Academic Press.