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Nucleic Acids Research, 1980, Vol. 8, No. 8 1823-1837
© 1980

Articles

Isolation and characterization of genomic DNA coding for {alpha}2 Type I collagen*

G. Vogeli, E.V. Avvedimento, M. Sullivan**, J.V. Maizel, Jr.**, G. Lozano, S.L. Adams, I. Pastan and B. de Crombrugghe

Laboratory of Molecular Biology NCI, NIH, Bethesda, MD 20205, USA **Laboratory of Molecular Genetics NICHD, NIH, Bethesda, MD 20205, USA

Received January 30, 1980.

We have isolated and characterized a segment of the chick {alpha}2 collagen gene by screening a library of chick genomic fragments using as hybridization probe an {alpha}2 collagen cDNA clone. Several clones were isolated and one of them, {lambda}gCOL 204, was used for further studies. The DNA of {lambda}gCOL 204 hybridizes to a unique species of mRNA the size of {alpha}2 collagen mRNA. This mRNA can be translated into a unique polypeptide which comigrates in SDS-gel electro-phoresis with pro-{alpha}2 collagen. Electron microscopic analysis by R-loop technique indicates that {lambda}gCOL 204 contains 7Kb of the {alpha}2 collagen gene. This 7 Kb piece constitutes the 3' end of the gene. The same clone also contains 9 Kb of DNA that is immediately adjacent to the 3' end of the {alpha}2 collagen gene. The cloned segment of the {alpha}2 collagen gene is interrupted by 8 intervening sequences of various lengths. The coding sequences for collagen in this clone add up to approximately 1,800 bp, which correspond to about 1/3 of {alpha}2 collagen mRNA. DNA sequence analysis of a small coding segment of XgCOL 204 reveals a characteristic collagen type sequence which encodes for an amino acid sequence identical to a sequence found in calf {alpha}2 collagen. The sequence of this region of the protein has not yet been determined for the chick {alpha}2 collagen.

*Abbreviations

{alpha}2 collagen, {alpha}2 type I collagen; SSC, 0.15M NaCl, 0.015 M Na citrate; DBM, diazobenzyloxymethyl; bp, base pairs; SDS, sodium dodecyl sulfate; Kb, kilo base pairs.


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