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MHC class I typing in a songbird with numerous loci and high polymorphism using motif-specific PCR and DGGE

Westerdahl, Helena LU ; Wittzell, Håkan LU ; von Schantz, Torbjörn LU and Bensch, Staffan LU (2004) In Heredity 92(6). p.534-542
Abstract
The major histocompatibility complex (MHC) has a central role in the specific immune defence of vertebrates. Exon 3 of MHC class I genes encodes the domain that binds and presents peptides from pathogens that trigger immune reactions. Here we develop a fast population screening method for detecting genetic variation in the MHC class I genes of birds. We found evidence of at least 15 exon 3 sequences in the investigated great reed warbler individual. The organisation of the great reed warbler MHC class I genes suggested that a locus-specific screening protocol is impractical due to the high similarity between alleles across loci, including the introns flanking exon 3. Therefore, we used motif-specific PCR to amplify two subsets of alleles (... (More)
The major histocompatibility complex (MHC) has a central role in the specific immune defence of vertebrates. Exon 3 of MHC class I genes encodes the domain that binds and presents peptides from pathogens that trigger immune reactions. Here we develop a fast population screening method for detecting genetic variation in the MHC class I genes of birds. We found evidence of at least 15 exon 3 sequences in the investigated great reed warbler individual. The organisation of the great reed warbler MHC class I genes suggested that a locus-specific screening protocol is impractical due to the high similarity between alleles across loci, including the introns flanking exon 3. Therefore, we used motif-specific PCR to amplify two subsets of alleles ( exon 3 sequences) that were separated with by DGGE. The motif-specific primers amplify a substantial proportion of the transcribed class I alleles (2-12 alleles per individual) from as many as six class I loci. Although not exhaustive, this gives a reliable estimate of the class I variation. The method is highly repeatable and more sensitive in detecting genetic variation than the RFLP method. The motif-specific primers also allow us to avoid screening pseudogenes. In our study population of great reed warblers, we found a high level of genetic variation in MHC class I, and no less than 234 DGGE genotypes were detected among 248 screened individuals. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Heredity
volume
92
issue
6
pages
534 - 542
publisher
Macmillan
external identifiers
  • wos:000221640500009
  • pmid:15162116
  • scopus:3042698704
ISSN
1365-2540
DOI
10.1038/sj.hdy.6800450
language
English
LU publication?
yes
id
ea5803c0-7e2a-49a5-85c1-71f8864ec448 (old id 136986)
date added to LUP
2007-06-25 14:09:17
date last changed
2017-08-06 03:48:27
@article{ea5803c0-7e2a-49a5-85c1-71f8864ec448,
  abstract     = {The major histocompatibility complex (MHC) has a central role in the specific immune defence of vertebrates. Exon 3 of MHC class I genes encodes the domain that binds and presents peptides from pathogens that trigger immune reactions. Here we develop a fast population screening method for detecting genetic variation in the MHC class I genes of birds. We found evidence of at least 15 exon 3 sequences in the investigated great reed warbler individual. The organisation of the great reed warbler MHC class I genes suggested that a locus-specific screening protocol is impractical due to the high similarity between alleles across loci, including the introns flanking exon 3. Therefore, we used motif-specific PCR to amplify two subsets of alleles ( exon 3 sequences) that were separated with by DGGE. The motif-specific primers amplify a substantial proportion of the transcribed class I alleles (2-12 alleles per individual) from as many as six class I loci. Although not exhaustive, this gives a reliable estimate of the class I variation. The method is highly repeatable and more sensitive in detecting genetic variation than the RFLP method. The motif-specific primers also allow us to avoid screening pseudogenes. In our study population of great reed warblers, we found a high level of genetic variation in MHC class I, and no less than 234 DGGE genotypes were detected among 248 screened individuals.},
  author       = {Westerdahl, Helena and Wittzell, Håkan and von Schantz, Torbjörn and Bensch, Staffan},
  issn         = {1365-2540},
  language     = {eng},
  number       = {6},
  pages        = {534--542},
  publisher    = {Macmillan},
  series       = {Heredity},
  title        = {MHC class I typing in a songbird with numerous loci and high polymorphism using motif-specific PCR and DGGE},
  url          = {http://dx.doi.org/10.1038/sj.hdy.6800450},
  volume       = {92},
  year         = {2004},
}