Single strand conformation polymorphism (SSCP) analysis for the detection of mutations in the CYP11B1 gene

• Published in: The journal of clinical endocrinology and metabolism 1996-06, Vol.81 (6), p.2389-2393
• Main Authors: Skinner, C A, Rumsby, G, Honour, J W
• Format: Article
• Language: English
• Subjects: Base Sequence; Child, Preschool; Cytochrome P-450 CYP11B2 - genetics; Exons; Female; Gene Deletion; Genes; Humans; Infant; Infant, Newborn; Male; Molecular Sequence Data; Multigene Family; Mutation; Oligonucleotide Probes - genetics; Polymorphism, Single-Stranded Conformational
• ISSN: 0021-972X
• DOI: 10.1210/jc.81.6.2389

The adrenal 11 beta-hydroxylase is a mitochondrial P450 enzyme encoded by the CYP11B1 gene, which is situated on chromosome 8q22 in tandem with the gene for aldosterone synthase (CYP11B2). Deficiency of 11 beta-hydroxylase results in the inability to convert 11-deoxycortisol to cortisol and accounts for 5-8% of cases of congenital adrenal hyperplasia. In the following study the CYP11B1 genes from eight individuals with 11 beta-hydroxylase deficiency were screened for mutations using single strand conformation polymorphism (SSCP) analysis. Sequence analysis of variant exons revealed a 28 bp deletion and a 5 bp duplication exon 2 and five missense mutations, G267R, G267D, Q356X, R427H and C494F, distributed throughout the gene. One of these mutations, G267R, and a G to A transversion at the third nucleotide position of codon 318 occur at the +1 position of the splice donor sites. Mutations were neither the result of gene conversion nor nonhomologous recombination between the two closely related CYP11B genes.