PARTIAL DNA-SEQUENCES DERIVED FROM 2 PREVIOUSLY UNKNOWN HLA CLASS-I GENES

Genomic DNA of HLA class I gene segments was amplified by the polymerase chain reaction (PCR). The amplified 0.8 kb gene segment encompassed sequences corresponding to an exon 2 (alpha-1 domain), an intron 2 and an exon 3 (alpha-2 domain). The PCR product was cloned in M13 and sequenced. Two previously unknown sequences were found. One of them (DAN2) had an open reading frame and intact intron splice sites, but we did not find evidence for transcription. Best homology (87.5%) was found with HLA-BeWo C.1, a recently described HLA-C gene. The other sequence (DAN4) is derived from a pseudogene, because the putative 3' splice site of intron 2 was changed from AG to ATG and the sequence corresponding to exon 3 had a shift in reading frame resulting in a stop codon.