ANALYSIS OF CFTR TRANSCRIPTS IN NASAL EPITHELIAL-CELLS AND LYMPHOBLASTS OF A CYSTIC-FIBROSIS PATIENT WITH 621+1G-]T AND 711+1G-]T MUTATIONS

被引:26
作者
ZIELENSKI, J
BOZON, D
MARKIEWICZ, D
AUBIN, G
SIMARD, F
ROMMENS, JM
TSUI, LC
机构
[1] HOSP SICK CHILDREN,DEPT GENET,555 UNIV AVE,TORONTO M5G 1X8,ONTARIO,CANADA
[2] UNIV TORONTO,DEPT MOLEC & MED GENET,TORONTO M5S 1A8,ONTARIO,CANADA
[3] HOP DEBROUSSE,SERV BIOCHIM,F-69322 LYON 05,FRANCE
[4] HOP CHICOUTIMI,FIBROSE KYST CLIN,QUEBEC CITY,PQ,CANADA
关键词
D O I
10.1093/hmg/2.6.683
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have analyzed the CFTR mRNA populations in a cystic fibrosis patient heterozygous for the 621+1G-->T and 711+1G-->T 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+1G-->T or 711+1G-->T alleles, respectively. The third product was apparently derived from an alternatively spliced mRNA species in the absence of the nominal splice site (in 621+1G-->T) 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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收藏
页码:683 / 687
页数:5
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