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Nucleic Acids Research, 1985, Vol. 13, No. 7 2207-2225
© 1985

Articles

Isolation and partial characterization of the entire human pro{alpha} I(II) collagen gene

Frank O. Sangiorgi, Virginia Benson-Chanda, Wouter J. de Wet*, Mark E. Sobel§, Petros Tsipouras+ and Francesco Ramirez

Departments of Obstetrics and Gynecology, University of Medicine and Dentistry of New Jersey-Rutgers Medical School Piscataway, NJ 08854, USA *Department of Biochemistry, Potchefstroom University Potchefstroom, 2520 South Africa §Laboratory of Pathology, National Institutes of Health Bethesda, MD 20205, USA +Pediatrics, University of Medicine and Dentistry of New Jersey-Rutgers Medical School Piscataway, NJ 08854, USA

Received February 4, 1985. Accepted March 7, 1985.

Using a cDNA probe specific for the bovine Type II procollagen, a series of overlapping genomic clones containing 45 kb of contiguous human DNA have been isolated. Sequencing of a 54 bp exon, number 29, provided direct evidence that the recombinant clones bear human Type II collagen sequences. Localization of the 5' and 3' ends of the gene indicated that the human Type II collagen gene is 30 kb in size. This value is significantly higher than that of the homologous avian gene. The segregation of a polymorphic restriction site in informative families conclusively demonstrated that the Type II gene is found in a single copy in the human haploid genome. Finally, sequencing of a triple helical domain exon has confirmed that a rearrangement leading to the fusion of two exons occurred in the pro{alpha}l(I) gene, following the divergence of the fibrillar collagens.


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