Direct sequencing of a HLA-DRB gene by polymerase chain reaction: Sequence variation in DRw8 specificity

Abstract

SummaryThe nucleotide sequence of a HLA-DRB gene with a predominant subtype of DRw8 specificity in Japanese (DRS.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 DRwS/DwS.1 and DRwS/Dw8.2 sequences: isoleucine67(ATC) instead of phenylalanine87(TTC) in DRwS/DwS.1 and serineSr(AGC) instead of aspartic acidaT(GAT) in DRw8/Dw8.2. On the other hand the DR8.1 and DRwS/DwS.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 DRwS/Dw8.3. Furthermore, an oligonucleotide probe specific for this sequence was synthesized and hybridized with 33 HLAtyped controls. This probe clearly distinguished the particular subtype from other DRw8 subtypes and specificities.