| Summary: | The aim of this project was to identify and characterise the AR gene defect responsible for AIS in a cohort of fourteen patients with the complete form of the syndrome. The chosen method for mutation detection was fluorescent semi-automated sequence analysis of the entire coding region of the gene. One of the reasons for choosing this technique was to determine the efficiency and practicality of this method of mutation detection compared to single strand conformation polymorphism (SSCP). Sequence analysis identified nine mutations, four of which were novel. Mutation information allowed the determination of carrier status in other family members in six cases. In patients for whom no mutations in the coding region of the gene were identified, the AR mRNA was analysed. Two novel mutations were identified which affect the AR mRNA transcript; one was the result of a duplication of exon B and the second was a splicing mutation affecting the branch point consensus sequence in intron 1. The results of this study have provided further evidence that all cases of CAIS are due to defects in the AR gene. In a separate part of the project, four mutations were introduced into an AR expression vector by <I>in vitro</I> mutagenesis and the effect of the mutations on the ability of the receptor to bind androgen and to active transcription were studied. In each case these functional studies confirmed that the mutations were responsible for the AR defect observed in the patients.
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