Development and optimization of a hybridization technique to type the classical class I and class II B genes of the chicken MHC.

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Potts, Nicola D 
Bichet, Coraline 
Merat, Laurence 
Guitton, Edouard 
Krupa, Andrew P 

The classical class I and class II molecules of the major histocompatibility complex (MHC) play crucial roles in immune responses to infectious pathogens and vaccines as well as being important for autoimmunity, allergy, cancer and reproduction. These classical MHC genes are the most polymorphic known, with roughly 10,000 alleles in humans. In chickens, the MHC (also known as the BF-BL region) determines decisive resistance and susceptibility to infectious pathogens, but relatively few MHC alleles and haplotypes have been described in any detail. We describe a typing protocol for classical chicken class I (BF) and class II B (BLB) genes based on a hybridization method called reference strand-mediated conformational analysis (RSCA). We optimize the various steps, validate the analysis using well-characterized chicken MHC haplotypes, apply the system to type some experimental lines and discover a new chicken class I allele. This work establishes a basis for typing the MHC genes of chickens worldwide and provides an opportunity to correlate with microsatellite and with single nucleotide polymorphism (SNP) typing for approaches involving imputation.

Avian, BF1, BF2, BLB1, BLB2, Recombination, Animals, Chickens, Genes, MHC Class I, Genes, MHC Class II, Nucleic Acid Hybridization, Polymorphism, Genetic, Polymorphism, Single-Stranded Conformational, Reference Standards, Sequence Analysis, DNA
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Springer Science and Business Media LLC
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Biotechnology and Biological Sciences Research Council (BB/H531319/1)
Wellcome Trust (089305/Z/09/Z)
Wellcome Trust (110106/Z/15/Z)
Biotechnology and Biological Sciences Research Council (BBSRC) CASE studentship Welcome Trust programme grant (089305/Z/09/Z)