A donor splice site mutation (1811+1G→C) in intron 11 of the CFTR gene identified in a patient of Macedonian origin

Splicing defects in the CFTR gene: Minigene analysis of two mutations, 1811+1G>C and 1898+3A>G

“…The 1811+1GNC mutation has been previously described by Petreska et al [2]. This mutation was either associated with the deltaF508 (18/204), with non-deltaF508 (82/204) or with normal (104/204) chromosomes.…”

Molecular basis of cystic fibrosis in the Republic of Macedonia

“…PCR primers and allele-specific oligonucleotides (ASOs) for specific mutations were either gifts from several laboratories or were synthesized on DNA Synthesizer (System 1, Beckman, Palo Alto, CA). The sequences of primer sets for amplification and subsequent single-strand conformational polymorphism (SSCP) and/or digestion of exons 2, 3, 4, 5, 6a, 8, 10, 11, 13, 15, 16, 17b, 18, 23; exon 7; and for denaturing gradient gel electrophoresis (DGGE) of exon 11, respectively, are given elsewhere (17)(18)(19). Additional primers that were used were: 5'-TI'CTCAGGGTATTTTAT-GAGAA-3' and 5'XATTAAAACATGTACGAT-ACAG-3' for PCR/SSCP of exon 4, 5'-GTGAATCGATGTGGTGACCA-3' and 5'-CTATGATGGGACAGTCT-3' for PCR/SSCP of exon 12, 5'-GTGAAATTGTCTGCCATCTTA-204 3' and 5'-TTCACTTACTGAACACAGTCTA-3' for PCR/SSCP and digestion of exon 19; 5'-TGAACCTGATGACACACTCA-3' and 5'-TCT-TCTTCGTTAATTTCTTC-3' for PCR of a 175 bp of exon 13, 5'-GAAGTACAATACTGAAT-TATG-3', 5'-TGACGTACAAGTATCAAATA-3', 5'-ATGGCATGGTACCTATAT-3', 5'-GCAGT-AAAAAATATAAAT-3' for PCRISSCP and sequencing of exon 20 and 5'-CAATA-CAATAAGGGAAAAAT-3', 5'-AATGATGTCA-GCTATATCAG-3', 5'-AAAAAGTTATTTAAG-TTA-3', 5'-CCATTTGTGTTGGTATGA-3' for PCR/SSCP and sequencing of exon 21.…”

“…Exons 2,3,4. 5,6a,7,8,10,11,12,17b,18,19,20,21,23 and the flanking sequences were examined by radioactive SSCP (25). The samples were denatured and run on 19:l non-denaturing PAG (4"C/15 W) with variation in the percentage (5% if the amplification products were shorter than 400 bp and 4% if they were longer than 400 bp).…”

“…The molecular characterization of the defects underlying CF in patients from the Republic of Macedonia resulted in determination of 58.4% (97/ 166) of the CF chromosomes. Table 1 summarizes the data for the different CFTR gene mutations, with the corresponding frequencies in the total number and in the particular ethnic group, haplotype association, and method of detection (3,19,22,23,(30)(31)(32)(33). AF508 mutation was present in 47.6% (n = 76/166) of the CF chromosomes.…”