Genotyping of feline MHC (FLA) class II DRB by PCR-RFLP method using group-specific primers.
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 I 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.
Kuwahara
Y
Y
Laboratory of Internal Medicine, Division of Veterinary Medicine, Faculty of Agriculture, Gifu University, Japan.
Kitoh
K
K
Kobayashi
R
R
Iwata
J
J
Ohne
R
R
Hosokawa-Kanai
T
T
Matsumoto
Y
Y
Kitagawa
H
H
Sasaki
Y
Y
D000818:Animals; D001483:Base Sequence; D002415:Cats / Q000235:genetics* / Q000276:immunology; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D017931:DNA Primers / Q000737:chemistry; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D005260:Female; D005802:Genes, MHC Class II / Q000235:genetics*; D005838:Genotype; D006684:HLA-DR Antigens / Q000737:chemistry / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D006684:HLA-DR Antigens; D004247:DNA