Isolation and characterization of a 15-kilobase genomic sequence coding for part of the Pro alpha 2 chain of sheep type I collagen.

C. D. Boyd, P. Tolstoshev, M. P. Schafer, B. C. Trapnell, H. C. Coon, P. J. Kretschmer, A. W. Nienhuis, Ronald Crystal

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

DNA fragments, prepared by partial Eco RI digestion of fetal sheep liver genomic DNA, were used to prepare a "library" of amplified genomic sequences with the lambda vector Charon 4A. Several recombinant plaques were identified by their ability to hybridize to 32P-labeled cDNA prepared from fetal sheep tendon type I procollagen mRNA. Two of these recombinant DNA bacteriophages (SpC3 and SpC7) were identified as containing procollagen pro alpha 2 gene sequences by their ability to specifically anneal to procollagen pro alpha 2 mRNA. Restriction endonuclease and hybridization to a cloned pro alpha 2 cDNA demonstrated that approximately half (2.5 kilobases) of the pro alpha 2 mRNA sequence is distributed over 15 kilobases of genomic DNA. Restriction maps of SpC3 and SpC7 demonstrated that these two DNA fragments contain overlapping sequences of the pro alpha 2 gene. Electron microscopy and R-loop analysis of SpC3 revealed that at least 12 to 16 intervening sequences are distributed throughout the length of this gene fragment.

Original languageEnglish
Pages (from-to)3212-3220
Number of pages9
JournalJournal of Biological Chemistry
Volume255
Issue number7
Publication statusPublished - 10 Apr 1980
Externally publishedYes

Fingerprint

Collagen Type I
Sheep
Procollagen
Genes
DNA
Messenger RNA
Pluto
Complementary DNA
Deoxyribonuclease EcoRI
Bacteriophages
Recombinant DNA
Genomic Library
Tendons
DNA Restriction Enzymes
Liver
Introns
Electron microscopy
Digestion
Electron Microscopy

ASJC Scopus subject areas

  • Biochemistry

Cite this

Boyd, C. D., Tolstoshev, P., Schafer, M. P., Trapnell, B. C., Coon, H. C., Kretschmer, P. J., ... Crystal, R. (1980). Isolation and characterization of a 15-kilobase genomic sequence coding for part of the Pro alpha 2 chain of sheep type I collagen. Journal of Biological Chemistry, 255(7), 3212-3220.

Isolation and characterization of a 15-kilobase genomic sequence coding for part of the Pro alpha 2 chain of sheep type I collagen. / Boyd, C. D.; Tolstoshev, P.; Schafer, M. P.; Trapnell, B. C.; Coon, H. C.; Kretschmer, P. J.; Nienhuis, A. W.; Crystal, Ronald.

In: Journal of Biological Chemistry, Vol. 255, No. 7, 10.04.1980, p. 3212-3220.

Research output: Contribution to journalArticle

Boyd, CD, Tolstoshev, P, Schafer, MP, Trapnell, BC, Coon, HC, Kretschmer, PJ, Nienhuis, AW & Crystal, R 1980, 'Isolation and characterization of a 15-kilobase genomic sequence coding for part of the Pro alpha 2 chain of sheep type I collagen.', Journal of Biological Chemistry, vol. 255, no. 7, pp. 3212-3220.
Boyd CD, Tolstoshev P, Schafer MP, Trapnell BC, Coon HC, Kretschmer PJ et al. Isolation and characterization of a 15-kilobase genomic sequence coding for part of the Pro alpha 2 chain of sheep type I collagen. Journal of Biological Chemistry. 1980 Apr 10;255(7):3212-3220.
Boyd, C. D. ; Tolstoshev, P. ; Schafer, M. P. ; Trapnell, B. C. ; Coon, H. C. ; Kretschmer, P. J. ; Nienhuis, A. W. ; Crystal, Ronald. / Isolation and characterization of a 15-kilobase genomic sequence coding for part of the Pro alpha 2 chain of sheep type I collagen. In: Journal of Biological Chemistry. 1980 ; Vol. 255, No. 7. pp. 3212-3220.
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