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Immunogenetics

, Volume 33, Issue 4, pp 247–254 | Cite as

Identification of expressed bovine class I MHC genes at two loci and demonstration of physical linkage

  • Albert Bensaid
  • Anita Kaushal
  • Cynthia L. Baldwin
  • Hans Clevers
  • John R. Young
  • Stephen J. Kemp
  • Niall D. MacHugh
  • Philip G. Toye
  • Alan J. Teale
Original Articles

Abstract

A cDNA library prepared from lymphocytes of a cow (E98), homozygous at major histocompatibility complex (MHC) loci (BoLA phenotype w10, KN104), was screened with a bovine MHC class I probe. Of the cDNA clones isolated, two, (2.1 and 5.1) were selected and showed divergence at both 5′ and 3′ termini. E98 DNA was digested with rare-cutter enzymes (Sfi I, Mlu I, Not I, and Cla I) and fragments were size-separated by field inversion gel electrophoresis (FIGE). Hybridization with an entire class I cDNA probe revealed multiple fragments generated by each enzyme. When the 3′ untranslated regions (UT) of 2.1 and 5.1 were used as probes, only one fragment was revealed in each digested sample, showing locus specificity of these probes in cattle. Further, DNA of transfected mouse fibroblasts L4 (expressing KN104) and L10 (expressing w10) hybridized to the 3UT regions of clones 2.1 and 5.1, respectively, Northern blot analysis of the mRNA of the L4 and L10 transfected cells provided further evidence that the cDNA clones 2.1 and 5.1 code for the BoLA-KN104 and BoLA-w10 class I molecules respectively, and thus these represent the products of two different genes. A long range physical mapping of the BoLA-w10 and KN104 genes was performed using FIGE analysis of DNA of and homozygous and an heterozygous animal. This analysis revealed that the BoLA-w10 and KN104 genes are separated by not more than 210 kilobases (kb) and that they are components of a multigene family spanning 1550 kb. As the] w10 gene is at the BoLA-A locus we assign the KN104 gene to a B locus.

Keywords

Major Histocompatibility Complex cDNA Clone Major Histocompatibility Complex Class Northern Blot Analysis Multigene Family 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Albert Bensaid
    • 1
  • Anita Kaushal
    • 1
  • Cynthia L. Baldwin
    • 1
  • Hans Clevers
    • 1
  • John R. Young
    • 1
  • Stephen J. Kemp
    • 1
  • Niall D. MacHugh
    • 1
  • Philip G. Toye
    • 1
  • Alan J. Teale
    • 1
  1. 1.International Laboratory for Research on Animal DiseasesNairobiKenya

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