Cloning and characterization of five overlapping cDNAs specific for the human proα 1(I) collagen chain

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Abstract
We report the cloning of five overlapping cDNAs bearing sequences specific for the human proα1(I) collagen chain. Poly-A RNA enriched for collagen sequences was purified from normal human fibroblasts and used as template to synthesize double stranded cDNA. The cDNA was inserted into the Eco RI site of pBR 322 by blunt-ending and dG:dC tailing. The clones were screened by colony hybridization using the original RNA population and the resulting five positive clones subjected to restriction endo-nuclease mapping analysis and DNA sequencing. These overlapping clones cover from residue 247 in the α chain to part of the 3′ end untranslated region of the proα1(I) mRNA for a total of 3400 nucleotides.