High-resolution typing for chicken BF2 (MHC class I) alleles by automated sequencing.
Sequence-based typing (SBT) was developed for major histocompatibility complex (MHC) class I and class II alleles in humans. We report here the development and application of a SBT method for alleles of the chicken BF2 locus (the more polymorphic of the two MHC class I loci in chickens). Exon 2 of the BF2 gene was selectively amplified from genomic DNA using a BF2 locus-specific PCR primer. Exon 2 sequences were sufficient to identify the 21 distinct BF2 alleles described in standard B haplotypes of Leghorns and in commercial broiler-breeder lines. Sixty-six samples from MHC typed, pedigreed chickens were tested, including 50 different heterozygous combinations. BF2 sequences from all B homozygotes were successfully amplified, and all combinations of BF2 alleles in heterozygotes were co-amplified equally. The two different BF2 alleles in heterozygotes could be identified unambiguously by distinct sequence motif patterns. In tests of samples of unknown B genotype in commercial broiler-breeder flocks, we identified expected BF2 alleles as well as an allele not previously encountered in one of the lines.
Livant
E J
EJ
Department of Poultry Science, Auburn University, Auburn, AL 36849, USA.
Ewald
S J
SJ
D000483:Alleles; D000818:Animals; D001947:Breeding / Q000379:methods; D002645:Chickens / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D005805:Genes, MHC Class I / Q000235:genetics*; D005838:Genotype; D017422:Sequence Analysis, DNA / Q000379:methods*
D017931:DNA Primers