The Turkey Major Histocompatibility Complex: Identification of Class II Genotypes by Restriction Fragment Length Polymorphism Analysis of Deoxyribonucleic Acid

Abstract

Using a chicken Class II MHC clone in Northern blot analysis, tissue-specific expression of turkey Class II MHC genes was observed in the embryonic bursa of Fabricius as well as in the adult spleen. In contrast, there was no detectable expression in the embryonic liver, brain, or spleen. Southern blot analysis of BamHI-digested turkey DNA revealed two restriction fragment length polymorphism (RFLP) patterns that did not deviate significantly from single-gene Mendelian inheritance. Further analysis of PvuII-digested DNA from 325 turkeys showed four distinct RFLP patterns that segregated within the turkey lines studied. Because the chicken Class II MHC clone hybridized specifically to mRNA in immune-associated tissues, and because it identified polymorphisms among turkeys, the chicken clone is suggested to identify four turkey Class II MHC genotypes. The current study provides good evidence that RFLP analysis of DNA can be used as a means for molecular genotyping at the MHC in turkeys.

The Turkey Major Histocompatibility Complex: Characterization by Mixed Lymphocyte, Graft-Versus-Host Splenomegaly, and Skin Graft Reactions

“…In a previous report, four turkey Class II MHC homozygous genotypes were identified by Southern blot analysis of DNA and hybridization with a chicken Class II MHC genomic probe (Emara et al, 1992). However, it was not determined whether these four genotypes were phenotypically distinct.…”

“…There 0(0) 0(0) 1 (0)* 3(3) ! Data are presented as relative splenomegaly indices (rSI) as described previously (Emara et al, 1992). An rSI > 1.3 indicates significant splenomegaly, whereas an rSI less than 1.3 indicates no splenomegaly.…”

“…A turkey subline segregating for specific MHC genotypes was established (Emara et al, 1992). As of the spring of 1991, this turkey line was in its fourth generation of selection based on MHC type.…”

“…A chicken MHC Class II genomic clone was used in Southern blot analysis to identify turkey MHC Class II genotypes (Emara et al, 1992). The chicken MHC Class II clone hybridized to turkey DNA under stringent conditions, and there was tissue-specific expression of the turkey Class II genes in the bursa of Fabricius and adult spleen.…”

A locus-wide approach to assessing variation in the avian MHC: the B-locus of the wild turkey

“…Southern hybridization methods have been used to estimate the number of genes and alleles/locus for the class I and II loci thereby providing a rough estimate of overall genetic variation at the MHC (Emara et al, 1992;Westerdahl et al, 2000;Freeman-Gallant et al, 2002;Richardson and Westerdahl, 2003;Alcaide et al, 2008). PCR-based techniques such as reference strand-mediated conformational analysis, single strand conformation polymorphism and denaturing gradient gel electrophoresis (DGGE) are also effective in estimating the number of alleles, especially when locus-specific primers are available (Ramon et al, 1998;Goto et al, 2002;Westerdahl et al, 2004;Knapp, 2005;Alcaide et al, 2010).…”