© 1993 Oxford University Press
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Analysis of CFTR transcripts in nasal epithelial cells and lymphoblasts of a cystic fibrosis patient with 621 +1G
T and 711 +1GT mutations
1Department of Genetics, The Hospital for Sick Children Toronto, Ontario M5G 1XB, Canada 2Service de Biochimie, Håpital Debrousse F-69322 Lyon, France 3Clinique de Fibrose Kystique, Håpital de Chicoutimi Quebec 4Department of Molecular and Medical Genetics, University of Toronto Toronto, Ontario M5S 1A8, Canada
*To whom correspondence should be addressed at: Department of Genetics, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada
Received February 10, 1993; Revised April 7, 1993; Accepted April 7, 1993
We have analyzed the CFTR mRNA populations in a cystic fibrosis patient heterozygous for the 621 +1G
T and 711+1GT mutations. Total RNA isolated from the nasal epithelial cells and Epstein—Barr virus-transformed lymphoblasts derived from this patient was reversely transcribed and a region extending from exon 3 to exon 7 of the gene was amplified by the polymerase chain reaction and analyzed. Three abnormal products were identified, suggesting the presence of three aberrant transcripts, and their profiles were identical in both cell types. Two of the products were found to be missing either exon 4 or exon 5 as anticipated from the transcripts from the 621+1GT or 711+1GT alleles, respectively. The third product was apparently derived from an alternatively spliced mRNA species in the absence of the nominal splice site (in 621+1GT) through the use of a cryptic splice donor sequence (TT528/GTGAGG) within exon 4. Although reading frames appeared to be preserved in all three putative transcripts, significant portions of the presumed first and second transmembrane spans as well as the immediately following cytoplasmic domain would be deleted from the mutant CFTR polypeptides, if made. These observations are consistent with a loss of CFTR function in this cystic fibrosis patient.
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