Genotyping of feline MHC(FLA) class 2 DRB by PCR-RFLP method using group-specific primers

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  • KUWAHARA Yasuhito
    Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University
  • KITOH Katsuya
    Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University
  • KOBAYASHI Rie
    Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University
  • IWATA Junko
    Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University
  • OHNE Rieko
    Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University
  • HOSOKAWA-KANAI Tomoko
    Division of Bio-regulatory Systems, Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo Nippon Institute for Biological Science
  • MATSUMOTO Yoshitsugu
    Division of Bio-regulatory Systems, Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo
  • KITAGAWA Hitoshi
    Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University
  • SASAKI Yoshihide
    Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University

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  • Genotyping of Feline MHC (FLA) Class II DRB by PCR-RFLP Method Using Group-Specific Primers.

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For genotyping of feline major histocompatibility complex (FLA) class II DRB, the polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method using group-specific primers was tried. Sixty-six DRB genes were classified into 8 groups according to differences in the first 5' amino acid sequences. The group-specific primers were designed as forward ones, which were specific for 5' base sequences of genes in each group. Three to 7 appropriate restricted enzymes were selected by computer analysis for RFLP typing of the genes divided into each group. In 6 out of 9 cats, the results of DRB typed by direct sequence method agreed with results of the PCR-RFLP method using group-specific primers. In the other 3 cats, the number of genes amplified by group-specific primers was 1 or 2 more than those detected by direct sequence method. The direct sequence method in 9 cats identified 5 new FLA-DRB genes. The PCR-RFLP method using group-specific primers could divide 66 genes into 37 genes and 10 subgroups from the RFLP pattern. One to 6 genes in each cat, and a total of 203 genes and subgroups were detected in 68 domestic cats. The genes detected might be biased to the subgroup G1-1a (28.8%), DRB*0501 (10.3%), G1-2a (9.4%) and G6b (7.4%). The PCR-RFLP method using group-specific primers may be useful in typing FLA class II DRB.

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