C to T polymorphism in exon 33 of the COL3A1 gene.

Source and Description of Sequence Polymorphism: Nucleotide sequencing of type HI procollagen (COL3A1) cDNAs from several individuals revealed a nucleotide polymorphism that converted the codon for amino acid 581 of the triple helix from -GGC(C-allele) (1) to -GGT(T-allele) in some alleles. The PCR was carried out with primers below; cDNA products were 226 bp and genomic products were 326 bp in length. Digestion of PCR products from cDNA with Haelll yielded fragments of 9, 33, 36, 58 and 90 bp for the C-allele and fragments of 9, 36, 90 and 91 bp for the T-allele. Digestion of PCR products from genomic DNA yielded fragments of 33, 34, 35 and 224 bp for the C-allele and fragments of 34, 35 and 257 bp for the T-allele. AUele-specific oligomer (ASO) hybridization was used to study 50 unrelated individuals of the following origins: 23 US Americans, 11 Finnish, 6 Swedish, 4 Canadian, 2 British, 2 Dutch and 2 Japanese. Heterozygous individuals were present in all nationalities. All experiments were repeated at least three times with different PCR products. Oligomers Used to Amplify cDNA: 5' GAG AAA GAG GAG GTC TT 3' (SP10) 5' CCA GTT TCA CCT CTC TCA C 3' (SP31) Oligomers Used to Amplify Genomic DNA: 5' CAA CAC TCC TGG AAA GTA ATC G 3' (in IVS32) 5' AGT GCA GGA CTG TCC CAT ATG 3' (in IVS33) ASO Hybridization Probes: