Identification of a double mutation (D160V-K161E) in the p67phox gene of a chronic granulomatous disease patient

In neutrophils of a chronic granulomatous disease (CGD) patient with a lack of p67phox the mRNA for p67phox was present in normal amount and size. This mRNA was reverse transcribed, and the coding region was analyzed by single-strand conformation polymorphism analysis, Direct DNA sequencing allowed the identification of a A(479)-to-T and A(481)-to-G substitution in exon 5 of the p67phox gene resulting in a double nonconservative amino acid change (160)Lys-to-Glu and (161)Asp-to-Val (D160V-K161E), This defect was found in the genomic DNA of this patient in heterozygous state and does not correspond to those previously found in other cases of CGD lacking the p67phox. (C) 1997 Academic Press.