Isolation, Cloning, and Sequencing of Porcine Agouti Exon 2 (porAex2)

Authors

  • YAN WANG,

    1. Department of Biology/Microbiology, South Dakota State University, Brookings, South Dakota
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  • CARL A. WESTBY,

    Corresponding author
    1. Department of Biology/Microbiology, South Dakota State University, Brookings, South Dakota
      Address reprint requests, to Professor Carl A. Westby, Department of Biology and Microbiology, College of Agriculture and Biological Sciences, Northern Plains Biostress Laboratory, Box 2140D. SDSU, Brookings, SD 57007-2142 USA.
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  • MARCUS JOHANSEN,

    1. Department of Biology/Microbiology, South Dakota State University, Brookings, South Dakota
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  • DONALD M. MARSHALL,

    1. Department of Biology/Microbiology, South Dakota State University, Brookings, South Dakota
    2. Department of Animal Science, South Dakota State University, Brookings, South Dakota
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  • NELS GRANHOLM

    1. Department of Biology/Microbiology, South Dakota State University, Brookings, South Dakota
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Address reprint requests, to Professor Carl A. Westby, Department of Biology and Microbiology, College of Agriculture and Biological Sciences, Northern Plains Biostress Laboratory, Box 2140D. SDSU, Brookings, SD 57007-2142 USA.

Abstract

In order to isolate, clone, and sequence agouti exon 2 of the pig (Yorkshire), we used an interspecific hybridization strategy. Primers from the 5′ and 3′ borders of the known human agouti exon 2 sequence were used to amplify (PCR) pig agouti exon 2. Following Southern blotting using a human exon 2 internal primer to authenticate that our PCR amplified product was truly pig exon 2 (PorAex2), the fragment was cloned and sequenced. PorAex2 exhibits 79.1 and 75.7% DNA sequence and 85 and 74% deduced amino acid sequence homologies with human and mouse agouti exon 2 and agouti protein, respectively. With the isolation of PorAex2, we can now map, sequence, and clarify the modus operandi of the porcine agouti gene. The GenBank Accession number of PorAex 2 is AF018166.

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