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Description, Source and Method: To date, only one RFLP (1) has been reported in the gene for the proa 1(1) chain of type I procollagen (COL1A1). cDNA and genomic DNA was PCR amplified with a 5' primer (5'-AGACCAGGAATTCGGCTTCG-3') spanning the EcoRI site in the proa 1(1) C-propeptide and a 3 ' primer (5 '-TTGGATCCAAGGTTGAATGCACTTTTGG-3') directed to nucleotides +203 to +222 (+1 being used to note the first nucleotide after the stop codon) and containing an additional BamHI site. Twenty-five cycles of amplification were performed at 94°C (1 min), 52°C (1 min), and 72°C (1 min) using a commercial kit (Cetus/Perkin-Elmer), and a DNA thermocycler. After digestion with EcoRI and BamHI, the 265 base pair fragment was cloned into M13 and sequenced. The two alleles were distinguished by allele specific oligonucleotide hybridization (2). The oligonucleotide specific for the allele with a C had the sequence 5'-TGGCCA-3' and the oligonucleotide specific for the allele with a T had the sequence 5 '-TGGCTTTTGATTCA-3'. Polymorphism: The sequence polymorphism was at position +88 (+1 being used to note the first nucleotide after the stop codon) of the proal(I) chain mRNA. The nucleotide at +88 was either a C or a T (Fig. 1). Frequency: Unrelated individuals studied: chromosomes from seven individuals of one family, from three individuals of another family, and from eight unrelated individuals were studied. Five individuals were heterozygotes for either allele, eleven were homozygote for a T and two homozygotes for a C at position + 88. Mendelian Inheritance: The T and C sequence polymorphism was shown to be inherited in Mendelian manner in 7 individuals from 3 generations of one family and in 3 individuals from two generations of another family.

A sequence polymorphism in the 3′-nontranslated region of the proα1 chain of type I procollagen