Use of constant denaturant capillary electrophoresis of pooled blood samples to identify single-nucleotide polymorphisms in the genes (Scnn1a and Scnn1b) encoding the alpha and beta subunits of the epithelial sodium channel.

Détails

ID Serval
serval:BIB_F8EA0CA034AE
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Use of constant denaturant capillary electrophoresis of pooled blood samples to identify single-nucleotide polymorphisms in the genes (Scnn1a and Scnn1b) encoding the alpha and beta subunits of the epithelial sodium channel.
Périodique
Clinical Chemistry
Auteur(s)
Xue M.Z., Bonny O., Morgenthaler S., Bochud M., Mooser V., Thilly W.G., Schild L., Leong-Morgenthaler P.M.
ISSN
0009-9147 (Print)
ISSN-L
0009-9147
Statut éditorial
Publié
Date de publication
2002
Peer-reviewed
Oui
Volume
48
Numéro
5
Pages
718-728
Langue
anglais
Résumé
BACKGROUND: The epithelial sodium channel (ENaC) is composed of three homologous subunits: alpha, beta, and gamma. Mutations in the Scnn1b and Scnn1g genes, which encode the beta and the gamma subunits of ENaC, cause a severe form of hypertension (Liddle syndrome). The contribution of genetic variants within the Scnn1a gene, which codes for the alpha subunit, has not been investigated. METHODS: We screened for mutations in the COOH termini of the alpha and beta subunits of ENaC. Blood from 184 individuals from 31 families participating in a study on the genetics of hypertension were analyzed. Exons 13 of Scnn1a and Scnn1b, which encode the second transmembrane segment and the COOH termini of alpha- and beta-ENaC, respectively, were amplified from pooled DNA samples of members of each family by PCR. Constant denaturant capillary electrophoresis (CDCE) was used to detect mutations in PCR products of the pooled DNA samples. RESULTS: The detection limit of CDCE for ENaC variants was 1%, indicating that all members of any family or up to 100 individuals can be analyzed in one CDCE run. CDCE profiles of the COOH terminus of alpha-ENaC in pooled family members showed that the 31 families belonged to four groups and identified families with genetic variants. Using this approach, we analyzed 31 rather than 184 samples. Individual CDCE analysis of members from families with different pooled CDCE profiles revealed five genotypes containing 1853G-->T and 1987A-->G polymorphisms. The presence of the mutations was confirmed by DNA sequencing. For the COOH terminus of beta-ENaC, only one family showed a different CDCE profile. Two members of this family (n = 5) were heterozygous at 1781C-->T (T594M). CONCLUSION: CDCE rapidly detects point mutations in these candidate disease genes.
Mots-clé
Electrophoresis, Capillary, Epithelial Sodium Channel, Exons, Female, Humans, Male, Pedigree, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, Protein Subunits, Sodium Channels/blood, Sodium Channels/genetics
Pubmed
Web of science
Création de la notice
24/01/2008 13:55
Dernière modification de la notice
03/03/2018 22:53
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