Fast Screening Method For Polymorphisms in Exon 9 of the Catalase Gene. - Fast Screening Method For Polymorphisms in Exon 9 of the Catalase Gene. - Open Access Pub

We checked our simple screening technique for detection of the known polymorphism of rs769217, and the two acatalasemic mutations in exon 9 of the catalase gene. This fast and inexpensive method yielded better resolution than those of the standard SSCP. We suppose that the method detects the spontaneously formed single stranded DNAs. DOI10.14302/issn.2575-7881.jdrr-16-1375 High concentrations of hydrogen peroxide are toxic for human cells and tissues but an increasing body of evidence indicates that its low concentrations function in intracellular signaling 1, 2, 3. The enzyme catalase (EC 1.11.1.6, CAT Gene ID: 847, NM_001752.3, NP_001743.1)4 is the main regulator of hydrogen peroxide metabolism, especially in erythrocytes 5. It decomposes hydrogen peroxide into oxygen and water in a concentration dependent manner. Due to the unique structure of catalase 6, 7 the enzyme decomposes the high (toxic) concentration of hydrogen peroxide while it seems to be unaffected on its low (physiologic) concentration (“flood and gate” mechanism) Inherited human catalase deficiency(acatalasemia) is associated with disorders8, 9 such as diabetes mellitus10, 11, vitiligo12, dyslipidemia13 and abnormal erythrocyte metabolism14. Fifteen catalase gene (acatalasemic) mutations responsible for decreased catalase activity have been reported from Japan (one missense in exon 4, one splicing in intron 4), North America (one missense in exon 9), Austria (one missense in exon 3) and eleven from Hungary 9, 11. In exon 9 two acatalasemic mutations were reported one from North America (c. 1093C>T, p.Arg365Cys)15 and two from Hungary (c.1093C>T, p.Arg365Cys and c.1060G>A, p.Arg354His) 8, 9. Furthermore, one silent substitution in exon 9 (rs:769217, c.1058C>T, p.Asp389Asp) was suggested to vitiligo susceptibility12. These reports indicate the increased frequency of catalase gene polymorphisms/mutations in exon 9. The first detection of rs769214 polymorphism was reported by Wen JK. et al. from Japan 15. They used DNA sequencing while the other authors applied the simpler methods of SSCP 14, and RPLF with BstX1 enzyme 12, 16, 17, 18, 19. The standard version of the single strand conformation polymorphism requires the denaturation (heat, formamide or both) of double strand PCR fragments. This denaturation often leads to the formation of several intermediate strands and sometimes the single strands could not be separated easily by polyacrylamide electrophoresis. The RFLP requires digestion of PCR product by restriction enzyme (BstX1). Whereas researchers at the next generation sequencing