Subject(s)
Cystic Fibrosis/genetics , Electrolytes/analysis , Mutation , Sweat/chemistry , Base Sequence , Cystic Fibrosis/metabolism , DNA/analysis , Exons , Female , Humans , Middle Aged , Molecular Sequence DataABSTRACT
Cystic fibrosis (CF) is a recessive disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. We have identified in exon 7 two frameshift mutations, one caused by a two-nucleotide insertion and the other caused by a one-nucleotide deletion; these mutations--CF1154insTC and CF1213delT, respectively, are predicted to shift the reading frame of the protein and to introduce UAA(ochre) termination codons at residues 369 and 368.
Subject(s)
Cystic Fibrosis/genetics , Frameshift Mutation , Adolescent , Adult , Amino Acid Sequence , Base Sequence , DNA/genetics , Electrophoresis, Polyacrylamide Gel , Exons , Humans , Molecular Sequence Data , Polymerase Chain Reaction , Polymorphism, GeneticABSTRACT
We established a pancreatic adenocarcinoma cell line (CFPAC-1) from a patient with cystic fibrosis (CF) and assessed some of its properties. The cells show epithelial morphology and express cytokeratin and oncofetal antigens characteristic of pancreatic duct cells. Basal and stimulated levels of cAMP and cAMP-dependent protein kinase and the biophysical properties of single Cl- channels in CFPAC-1 are similar to those of airway and sweat gland primary cultures and Cl(-)-secreting epithelial cell lines. Anion transport and single Cl- channel activity was stimulated by Ca2+ ionophores but not by forskolin, cAMP analogs, or phosphodiesterase inhibitors. The cells express the CF gene and manifest the most common CF mutation, deletion of three nucleotides resulting in a phenylalanine-508 deletion. These properties have been stable through greater than 80 passages (24 months), suggesting that CFPAC-1 can serve as a continuous cell line that displays the CF defect.