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Deletion Pattern in the 21‐Hydroxylase Gene Detected by Polymerase Chain Reaction
Author(s) -
Kinoshita Eiichi,
Matsumoto Tadashi,
Kondoh Tatsuro,
Yoshimoto Masaaki,
Niikawa Norio,
Tsuji Yoshiro
Publication year - 1991
Publication title -
pediatrics international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.49
H-Index - 63
eISSN - 1442-200X
pISSN - 1328-8067
DOI - 10.1111/j.1442-200x.1991.tb01512.x
Subject(s) - pseudogene , polymerase chain reaction , microbiology and biotechnology , gene , taq polymerase , genetics , mutation , gene mutation , dna , medicine , biology , genome , thermus aquaticus
In order to detect deletion mutation and/or gene conversion in the 21‐hydroxylase (21‐OH) gene, we adopted the polymerase chain reaction (PCR) method followed by electrophoresis. Two pairs of synthesized primers, Ta/lb and 2a/2b, each corresponding to the sequence at the 5′ portion of the 21‐OH gene, were set for PCR. Taq l digestion of amplified DNA from normal individuals using Ta/lb as primers gave the following three fragments: an active 21‐OH gene‐derived 559 bp fragment, and pseudogene‐derived 364 and 195 bp fragments. Of 16 patients with 21‐hydroxylase deficiency (21‐OHD) studied, 6 (37%) lacked the 559 bp fragment. These 6 patients also lacked both the 331 and 117 bp Mval fragments of the PCR product which were obtained with the primers 2a/2b, both being derived from the active 21‐OH gene. These results indicate that 6 of the 16 patients have either deletion of the 21‐OH gene or conversion of the gene to its tandemly located pseudogene. The method described here provides a rapid diagnosis of 21‐OHD.