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Identification and phylogenetic analysis of 15 MHC class II DRB1 β1 expressed alleles in a ewe–lamb flock

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Abstract

Ovar-DRB1 is part of the major histocompatibility complex (MHC) class II of sheep and functions by presenting extracellular-derived peptides to the immune system. Although there are over 100 different Ovar-DRB1 DNA sequences reported in GenBank, only two Ovar-DRB1 mRNA sequences have been reported. As a first step in understanding MHC Class II function as it relates to disease progression in sheep, Ovar-DRB1 transcripts encoding the peptide-binding site or the first domain (β1) of Ovar-DRB1 in a 32-ewe–lamb flock were identified and characterized by using reverse transcriptase–polymerase chain reaction, cloning, sequencing, and phylogenetic analysis. Fourteen new Ovar-DRB1 β1 cDNA sequences out of a total of 15 Ovar-DRB1 β1 cDNA sequences in a ewe–lamb flock of 32 sheep were identified. One Ovar-DRB1 β1 cDNA sequence was 100% identical to M93432, one of the two Ovar-DRB1 mRNA sequences reported in GenBank. Twelve out of 15 Ovar-DRB1 β1 cDNA sequences were 100% identical to the corresponding previously reported Ovar-DRB1 genomic DNA sequences, indicating that these Ovar-DRB1 genomic DNA sequences are also transcribed. One of three of the remaining Ovar-DRB1 β1 cDNA sequences, DRB1*07012, had a synonymous substitution resulting in an identical deduced amino acid sequence to DRB1*0701. Two of the remaining three Ovar-DRB1 β1 cDNA sequences had nucleotide differences and subsequent deduced amino acid sequence differences when compared to known Ovar-DRB1 β1 genomic DNA sequences, and therefore, DRB1*0206 and DRB1*0353 represent new Ovar-DRB1 β1 expressed alleles. Phylogenetic analysis of the 15 Ovar-DRB1 β1 cDNA sequences revealed that DRB1*0206 had a strong phylogenetic relationship to DRB1*0203, and DRB1*0353 had a strong phylogenetic relationship to DRB1*0303.

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Acknowledgements

We thank Janet Alverson for the critical reading of this manuscript. We thank Nancy Kumpula-McWhirter, Nicholas Durfee, and Liam Broughton for their technical assistance. We also thank Emma Karel, Pete Steiner, Duane Chandler, Lori Fuller, and Dr. Steve Parish for animal care. This work was supported by USDA-ARS CWD #5348-32000-019-00D. All work performed in this manuscript involving live animal research was performed with approval from the Institutional Animal Care and Use Committee (IACUC) of Washington State University, Pullman, Washington, USA

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Authors and Affiliations

  1. Animal Disease Research Unit, USDA-ARS, 3003 ADBF, Washington State University, Pullman, WA, 99164-6630, USA

    Lynn M. Herrmann & Donald P. Knowles

  2. Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, 99164-7040, USA

    Lynn M. Herrmann, Wendy C. Brown & Donald P. Knowles

  3. US Sheep Experiment Station, Agricultural Research Service, US Department of Agriculture, Dubois, ID, 83423, USA

    Greg S. Lewis

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  1. Lynn M. Herrmann
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  2. Wendy C. Brown
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  3. Greg S. Lewis
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  4. Donald P. Knowles
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Correspondence to Lynn M. Herrmann.

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Herrmann, L.M., Brown, W.C., Lewis, G.S. et al. Identification and phylogenetic analysis of 15 MHC class II DRB1 β1 expressed alleles in a ewe–lamb flock. Immunogenetics 57, 855–863 (2005). https://doi.org/10.1007/s00251-005-0050-9

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  • DOI: https://doi.org/10.1007/s00251-005-0050-9

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