Low yield of polymorphisms from EST blast searching: analysis of genes related to oxidative stress and verification of the P197L polymorphism in GPX1.

摘要: To determine new polymorphisms in the antioxidant enzymes superoxide dismutase, glutathione peroxidases, catalase, and microsomal transferase 1, a search of human expressed sequence tags (EST) database was performed (with BLAST 2.0). When any mutation, indicated by search, gave rise to nonconservative amino acid change we polymerase chain reaction (PCR) restriction analysis and/or genomic DNA from subjects order verify these potential polymorphisms. Of nine EST found four different enzymes, could one, an substitution Pro-Leu at position 197 (P197L), peroxidase 1 gene. The corresponding allele frequencies were approximately 70/30%. In addition, silent mutation (1167T/C) catalase gene also be verified. Six individuals analyzed per polymorphism, so that only common would found. mutations not verified direct thus cannot excluded as allelic variation population. These results show can used for genes with high abundance database. addition analysis, PCR/single-strand conformation polymorphism (SSCP) employed No coding detected either method. degree conservation indicates important physiological function this enzyme.