Abstract
Molecular genetic analysis was performed in a patient with cytochrome b positive X-linked chronic granulomatous disease. A previous Southern blot study, using a cytochrome b heavy chain cDNA as probe, revealed a Pst I restriction fragment pattern for the cytochrome b heavy chain gene (CYBB) different to that of normal individuals. Since restriction length polymorphism with Pst I has never been observed in control individuals and no abnormal restriction fragment patterns in the patient's CYBB was detected with seven other enzymes used, we focussed on the single Pst I site in the CYBB cDNA as being the only mutation site responsible for his disease. A fragment of the patient's cDNA which included the Pst I site was amplified by reverse polymerase chain reaction, and loss of the Pst I site in the fragment was confirmed by incubation with Pst I. Subsequent sequence analysis of the fragment revealed a point mutation in the Pst I site (cytosine to adenine), substituting glutamic acid for alanine at position 57.
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Abbreviations
- CGD:
-
chronic granulomatous disease
- CYBB:
-
cytochrome b heavy chain gene
- EBV:
-
Epstein-Barr virus
- phox:
-
phagocytic oxidase
- gp91-phox:
-
91-kD glycoprotein
- p22-phox:
-
22-kD polypepetide
- p47-phox:
-
47-kD cytoplasmic oxidase component
- p67-phox:
-
67-kD cytoplasmic oxidase component
- PCR:
-
polymerase chain reaction
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Ariga, T., Sakiyama, Y., Tomizawa, K. et al. A newly recognized point mutation in the cytochrome b558 heavy chain gene replacing alanine57 by glutamic acid, in a patient with cytochrome b positive X-linked chronic granulomatous disease. Eur J Pediatr 152, 469–472 (1993). https://doi.org/10.1007/BF01955051
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DOI: https://doi.org/10.1007/BF01955051