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Evolutionary dynamics of an expressed MHC class IIβ beta locus in the Ranidae (Anura) uncovered by genome walking and high-throughput amplicon sequencing

Mulder, Kevin P. (author)
Smithsonian Conservat Biol Inst, Ctr Conservat Genom, Natl Zool Pk.;CIBIO InBIO, Res Ctr Biodivers & Genet Resources.
Cortazar-Chinarro, Maria (author)
Uppsala universitet,Zooekologi
Harris, D. James (author)
CIBIO InBIO, Res Ctr Biodivers & Genet Resources.
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Crottini, Angelica (author)
CIBIO InBIO, Res Ctr Biodivers & Genet Resources.
Grant, Evan H. Campbell (author)
US Geol Survey, Patuxent Wildlife Res Ctr, SO Conte Anadromous Fish Res Lab.
Fleischer, Robert C. (author)
Smithsonian Conservat Biol Inst, Ctr Conservat Genom, Natl Zool Pk.
Savage, Anna E. (author)
Smithsonian Conservat Biol Inst, Ctr Conservat Genom, Natl Zool Pk.;Univ Cent Florida, Dept Biol.
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Smithsonian Conservat Biol Inst, Ctr Conservat Genom, Natl Zool Pk;CIBIO InBIO, Res Ctr Biodivers & Genet Resources. Zooekologi (creator_code:org_t)
Oxford : Elsevier, 2017
2017
English.
In: Developmental and Comparative Immunology. - Oxford : Elsevier. - 0145-305X .- 1879-0089. ; 76, s. 177-188
  • Journal article (peer-reviewed)
Abstract Subject headings
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  • The Major Histocompatibility Complex (MHC) is a genomic region encoding immune loci that are important and frequently used markers in studies of adaptive genetic variation and "disease resistance. Given the primary role of infectious diseases in contributing to global amphibian declines, we characterized the hypervariable exon 2 and flanking introns of the MHC Class II beta chain for 17 species of frogs in the Ranidae, a speciose and cosmopolitan family facing widespread pathogen infections and declines. We find high levels of genetic variation concentrated in the Peptide Binding Region (PBR) of the exon. Ten codons are under positive selection, nine of which are located in the mammal-defined PBR. We hypothesize that the tenth codon (residue 21) is an amphibian-specific PBR site that may be important in disease resistance. Trans-species and trans-generic polymorphisms are evident from exon-based genealogies, and co-phylogenetic analyses between intron, exon and mitochondrial based reconstructions reveal incongruent topologies, likely due to different locus histories. We developed two sets of barcoded adapters that reliably amplify a single and likely functional locus in all screened species using both 454 and Illumina based sequencing methods. These primers provide a resource for multiplexing and directly sequencing hundreds of samples in a single sequencing run, avoiding the labour and chimeric sequences associated with cloning, and enabling MHC population genetic analyses. Although the primers are currently limited to the 17 species we tested, these sequences and protocols provide a useful genetic resource and can serve as a starting point for future disease, adaptation and conservation studies across a range of anuran taxa.

Subject headings

NATURVETENSKAP  -- Biologi -- Zoologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences -- Zoology (hsv//eng)
NATURVETENSKAP  -- Biologi -- Immunologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences -- Immunology (hsv//eng)

Keyword

MHC class II beta
Ranidae
Amplicon primers
Illumina
454
Lithobates
Rana
Pelophylax
Odorrana

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ref (subject category)
art (subject category)

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