Utility of three restriction fragment length polymorphism probes for genotyping of the chicken major histocompatibility complex class IV region

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Abstract
Three chicken B-G cDNA probes (gene 8.5, bg28, and bg32.1) were used to detect restriction fragment length polymorphisms (RFLP) in the chicken MHC class IV (B-G). By using inbred and selected chicken lines with different B haplotypes identified by hemagglutination, we identified B haplotypes (B2, B9, B11, B12, B15, B19, B21, B31, and B32) by RFLP using the three probes following digestion of genomic DNA with four restriction endonucleases (BglII, EcoRI, HaeIII, and PvuII). The GSP inbred line, previously shown to contain B-F21 by the use of a monoclonal antibody, did not contain B-G21, based on RFLP tests, whereas line N had B-F21 and B-G21. Consequently, the RFLP typing with the clone of B-G cDNA was able to determine the B haplotype in more detail than typing by hemagglutination. In inbred and selected lines, three B-G cDNA are useful DNA probes for RFLP to identify B genotypes. Two families of chickens with segregating B haplotypes were analyzed by RFLP using these probes; however, identification of the B genotype by this method was difficult in the randomly bred population. Genotypic comparisons of RFLP with gene 8.5 and BglII and bg 28 as probes and digestion by the endonucleases EcoRI, HaeIII, and PvuII between the parents and their offspring were generally compatible within the expectations of Mendelian inheritance.