A new approach for identifying non-pathogenic mutations. An analysis of the cystic fibrosis transmembrane regulator gene in normal individuals

Cristina Bombieri, Silvia Giorgi, Soukeyna Carles, Rafael De Cid, Francesca Belpinati, Caterina Tandoi, Nathalie Pallares-Ruiz, Conxi Lazaro, Bianca Maria Ciminelli, Marie Catherine Romey, Teresa Casals, Fiorenza Pompei, Giorgio Gandini, Mireille Claustres, Xavier P. Estivill, Pier Franco Pignatti, Guido Modiano

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Given q as the global frequency of the alleles causing a disease, any allele with a frequency higher than q minus the cumulative frequency of the previously known disease-causing mutations (threshold) cannot be the cause of that disease, This principle was applied to the analysis of cystic fibrosis transmembrane conductance regulator (CFTR) mutations in order to decide whether they are the cause of cystic fibrosis. A total of 191 DNA samples from random individuals from Italy, France, and Spain were investigated by DGGE (denaturing gradient gel electrophoresis) analysis of all the coding and proximal non-coding regions of the gene. The mutations detected by DGGE were identified by sequencing. The sample size was sufficient to select essentially all mutations with a frequency of at least 0.01. A total of 46 mutations was detected, 20 of which were missense mutations. Four new mutations were identified: 1341 + 28 C/T, 2082 C/T, L1096R, and I1131V. Thirteen mutations (125 G/C, 875 + 40 A/G, TTGAn, IVS8-6 5T, IVS8-6 9T, 1525-61 A/G, M470V, 2694 T/G, 3061-65 C/A, 4002 A/G, 4521 G/A, IVS8 TG10, IVS8 TG12) were classified as non-CF-causing alleles on the basis of their frequency. The remaining mutations have a cumulative frequency far exceeding q; therefore, most of them cannot be CF-causing mutations. This is the first random survey capable of detecting all the polymorphisms of the coding sequence of a gene.

Original languageEnglish
Pages (from-to)172-178
Number of pages7
JournalHuman Genetics
Volume106
Issue number2
DOIs
Publication statusPublished - 2000
Externally publishedYes

Fingerprint

Regulator Genes
Cystic Fibrosis
Mutation
Denaturing Gradient Gel Electrophoresis
Alleles
Cystic Fibrosis Transmembrane Conductance Regulator
Missense Mutation
Gene Frequency
Spain
Sample Size
Italy
Genes
France
DNA

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

A new approach for identifying non-pathogenic mutations. An analysis of the cystic fibrosis transmembrane regulator gene in normal individuals. / Bombieri, Cristina; Giorgi, Silvia; Carles, Soukeyna; De Cid, Rafael; Belpinati, Francesca; Tandoi, Caterina; Pallares-Ruiz, Nathalie; Lazaro, Conxi; Ciminelli, Bianca Maria; Romey, Marie Catherine; Casals, Teresa; Pompei, Fiorenza; Gandini, Giorgio; Claustres, Mireille; Estivill, Xavier P.; Pignatti, Pier Franco; Modiano, Guido.

In: Human Genetics, Vol. 106, No. 2, 2000, p. 172-178.

Research output: Contribution to journalArticle

Bombieri, C, Giorgi, S, Carles, S, De Cid, R, Belpinati, F, Tandoi, C, Pallares-Ruiz, N, Lazaro, C, Ciminelli, BM, Romey, MC, Casals, T, Pompei, F, Gandini, G, Claustres, M, Estivill, XP, Pignatti, PF & Modiano, G 2000, 'A new approach for identifying non-pathogenic mutations. An analysis of the cystic fibrosis transmembrane regulator gene in normal individuals', Human Genetics, vol. 106, no. 2, pp. 172-178. https://doi.org/10.1007/s004390051025
Bombieri, Cristina ; Giorgi, Silvia ; Carles, Soukeyna ; De Cid, Rafael ; Belpinati, Francesca ; Tandoi, Caterina ; Pallares-Ruiz, Nathalie ; Lazaro, Conxi ; Ciminelli, Bianca Maria ; Romey, Marie Catherine ; Casals, Teresa ; Pompei, Fiorenza ; Gandini, Giorgio ; Claustres, Mireille ; Estivill, Xavier P. ; Pignatti, Pier Franco ; Modiano, Guido. / A new approach for identifying non-pathogenic mutations. An analysis of the cystic fibrosis transmembrane regulator gene in normal individuals. In: Human Genetics. 2000 ; Vol. 106, No. 2. pp. 172-178.
@article{5a7925e6f9c448e5bac0fa0836089256,
title = "A new approach for identifying non-pathogenic mutations. An analysis of the cystic fibrosis transmembrane regulator gene in normal individuals",
abstract = "Given q as the global frequency of the alleles causing a disease, any allele with a frequency higher than q minus the cumulative frequency of the previously known disease-causing mutations (threshold) cannot be the cause of that disease, This principle was applied to the analysis of cystic fibrosis transmembrane conductance regulator (CFTR) mutations in order to decide whether they are the cause of cystic fibrosis. A total of 191 DNA samples from random individuals from Italy, France, and Spain were investigated by DGGE (denaturing gradient gel electrophoresis) analysis of all the coding and proximal non-coding regions of the gene. The mutations detected by DGGE were identified by sequencing. The sample size was sufficient to select essentially all mutations with a frequency of at least 0.01. A total of 46 mutations was detected, 20 of which were missense mutations. Four new mutations were identified: 1341 + 28 C/T, 2082 C/T, L1096R, and I1131V. Thirteen mutations (125 G/C, 875 + 40 A/G, TTGAn, IVS8-6 5T, IVS8-6 9T, 1525-61 A/G, M470V, 2694 T/G, 3061-65 C/A, 4002 A/G, 4521 G/A, IVS8 TG10, IVS8 TG12) were classified as non-CF-causing alleles on the basis of their frequency. The remaining mutations have a cumulative frequency far exceeding q; therefore, most of them cannot be CF-causing mutations. This is the first random survey capable of detecting all the polymorphisms of the coding sequence of a gene.",
author = "Cristina Bombieri and Silvia Giorgi and Soukeyna Carles and {De Cid}, Rafael and Francesca Belpinati and Caterina Tandoi and Nathalie Pallares-Ruiz and Conxi Lazaro and Ciminelli, {Bianca Maria} and Romey, {Marie Catherine} and Teresa Casals and Fiorenza Pompei and Giorgio Gandini and Mireille Claustres and Estivill, {Xavier P.} and Pignatti, {Pier Franco} and Guido Modiano",
year = "2000",
doi = "10.1007/s004390051025",
language = "English",
volume = "106",
pages = "172--178",
journal = "Human Genetics",
issn = "0340-6717",
publisher = "Springer Verlag",
number = "2",

}

TY - JOUR

T1 - A new approach for identifying non-pathogenic mutations. An analysis of the cystic fibrosis transmembrane regulator gene in normal individuals

AU - Bombieri, Cristina

AU - Giorgi, Silvia

AU - Carles, Soukeyna

AU - De Cid, Rafael

AU - Belpinati, Francesca

AU - Tandoi, Caterina

AU - Pallares-Ruiz, Nathalie

AU - Lazaro, Conxi

AU - Ciminelli, Bianca Maria

AU - Romey, Marie Catherine

AU - Casals, Teresa

AU - Pompei, Fiorenza

AU - Gandini, Giorgio

AU - Claustres, Mireille

AU - Estivill, Xavier P.

AU - Pignatti, Pier Franco

AU - Modiano, Guido

PY - 2000

Y1 - 2000

N2 - Given q as the global frequency of the alleles causing a disease, any allele with a frequency higher than q minus the cumulative frequency of the previously known disease-causing mutations (threshold) cannot be the cause of that disease, This principle was applied to the analysis of cystic fibrosis transmembrane conductance regulator (CFTR) mutations in order to decide whether they are the cause of cystic fibrosis. A total of 191 DNA samples from random individuals from Italy, France, and Spain were investigated by DGGE (denaturing gradient gel electrophoresis) analysis of all the coding and proximal non-coding regions of the gene. The mutations detected by DGGE were identified by sequencing. The sample size was sufficient to select essentially all mutations with a frequency of at least 0.01. A total of 46 mutations was detected, 20 of which were missense mutations. Four new mutations were identified: 1341 + 28 C/T, 2082 C/T, L1096R, and I1131V. Thirteen mutations (125 G/C, 875 + 40 A/G, TTGAn, IVS8-6 5T, IVS8-6 9T, 1525-61 A/G, M470V, 2694 T/G, 3061-65 C/A, 4002 A/G, 4521 G/A, IVS8 TG10, IVS8 TG12) were classified as non-CF-causing alleles on the basis of their frequency. The remaining mutations have a cumulative frequency far exceeding q; therefore, most of them cannot be CF-causing mutations. This is the first random survey capable of detecting all the polymorphisms of the coding sequence of a gene.

AB - Given q as the global frequency of the alleles causing a disease, any allele with a frequency higher than q minus the cumulative frequency of the previously known disease-causing mutations (threshold) cannot be the cause of that disease, This principle was applied to the analysis of cystic fibrosis transmembrane conductance regulator (CFTR) mutations in order to decide whether they are the cause of cystic fibrosis. A total of 191 DNA samples from random individuals from Italy, France, and Spain were investigated by DGGE (denaturing gradient gel electrophoresis) analysis of all the coding and proximal non-coding regions of the gene. The mutations detected by DGGE were identified by sequencing. The sample size was sufficient to select essentially all mutations with a frequency of at least 0.01. A total of 46 mutations was detected, 20 of which were missense mutations. Four new mutations were identified: 1341 + 28 C/T, 2082 C/T, L1096R, and I1131V. Thirteen mutations (125 G/C, 875 + 40 A/G, TTGAn, IVS8-6 5T, IVS8-6 9T, 1525-61 A/G, M470V, 2694 T/G, 3061-65 C/A, 4002 A/G, 4521 G/A, IVS8 TG10, IVS8 TG12) were classified as non-CF-causing alleles on the basis of their frequency. The remaining mutations have a cumulative frequency far exceeding q; therefore, most of them cannot be CF-causing mutations. This is the first random survey capable of detecting all the polymorphisms of the coding sequence of a gene.

UR - http://www.scopus.com/inward/record.url?scp=0034016792&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034016792&partnerID=8YFLogxK

U2 - 10.1007/s004390051025

DO - 10.1007/s004390051025

M3 - Article

VL - 106

SP - 172

EP - 178

JO - Human Genetics

JF - Human Genetics

SN - 0340-6717

IS - 2

ER -