Direct sequencing of a HLA-DRB gene by polymerase chain reaction: sequence variation in DRw8 specificity.
The nucleotide sequence of a HLA-DRB gene with a predominant subtype of DRw8 specificity in Japanese (DR8.1) was determined with single-stranded DNA enzymatically amplified by polymerase chain reaction (PCR). The sequence differs at a single amino acid from both of the published DRw8/Dw8.1 and DRw8/Dw8.2 sequences: isoleucine67(AUC) instead of phenylalanine67(TTC) in DRw8/Dw8.1 and serine57(AGC) instead of aspartic acid57(GAT) in DRw8/Dw8.2. On the other hand the DR8.1 and DRw8/Dw8.3 have the same amino acid sequence although one silent nucleotide substitution has occurred between the two sequences. These results indicate that Japanese DR8.1 specificity corresponds to DRw8/Dw8.3. Furthermore, an oligonucleotide probe specific for this sequence was synthesized and hybridized with 33 HLA-typed controls. This probe clearly distinguished the particular subtype from other DRw8 subtypes and specificities.
Department of Anthropology, Faculty of Science, University of Tokyo, Japan.
Y
Y
Watanabe
K
K
Tokunaga
K
K
Matsuki
K
K
Omoto
T
T
Juji
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D004277:DNA, Single-Stranded; D005007:Ethnology; D014644:Genetic Variation; D006684:HLA-DR Antigens / Q000235:genetics*; D059810:HLA-DR Serological Subtypes; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D004277:DNA, Single-Stranded; D006684:HLA-DR Antigens; D059810:HLA-DR Serological Subtypes; C041559:HLA-DR8 antigen