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Isolation and complete nucleotide sequence of the gene for bovine parathyroid hormone
Authors:C A Weaver  D F Gordon  M S Kissil  D A Mead  B Kemper
Affiliation:Department of Physiology and Biophysics and College of Medicine at Urbana-Champaign, University of Illinois at UrbanaChampaign, Urbana, IL 61801 U.S.A. Tel. (217)333-1146
Abstract:The structure of the bovine parathyroid hormone (PTH) gene has been analyzed by Southern blot hybridization of genomic DNA and by nucleotide sequence analysis of a cloned PTH gene. In the Southern analysis, several restriction enzymes produced single fragments that hybridized to PTH cDNA suggesting that there is a single bovine PTH gene. The restriction map of the cloned gene is the same as that determined by Southern blot analysis of bovine DNA. The sequence of 3154 bp of the cloned gene has been determined including 510 bp and 139 bp in the 5' and 3' flanking regions, respectively. The gene contains two introns which separate three exons that code primarily for: (i) the 5' untranslated region, (ii) the pre-sequence of preProPTH, and (iii) PTH and the 3' untranslated region. The gene contains 68% A + T and unusually long stretches of 100- to 150-bp sequences containing alternating A and T nucleotides in the 5' flanking region and intron A. The 5' flanking region contains two TATA sequences, both of which appear to be functional as determined by S1 nuclease mapping. Compared to the rat and human genes, the locations of the introns are identical but the sizes differ. Comparable human and bovine sequences in the flanking regions and introns are about 80% homologous.
Keywords:Recombinant DNA  Southern blot  S1 mapping  comparison with human gene  plasmid vector pBR322  M13 and λ phage  Charon 30  introns  exons  bp  base pairs  kb  1000 bp  pfu  plaque-forming units  preProPTH  preproparathyroid hormone  proPTH  proparathyroid hormone  PTH  parathyroid hormone  SDS  sodium dodecyl sulfate
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