Specific amplification of the HLA-DRB4 gene from c-DNA. Complete coding sequence of the HLA alleles DRB4*0103101 and DRB4*01033.

We present the complete coding sequence of the HLA alleles DRB40103101 and DRB401033 derived from the lymphoblastoid cell line G081, established from an individual of Spanish Gypsy ethnic origin. This cell was typed by PCR-SSP and reverse SSO as DRB40103101 but further characterization of the DRB4 gene by sequence-based typing (SBT) demonstrated heterozygosity at codon 78 (TAC, TAT). With the aim of confirming this polymorphism, RNA isolated from G081 was subjected to RT-PCR using primers designed to recognize specifically the 5' and 3' UT regions of HLA-DRB4 and the product was cloned and sequenced. Nucleotide sequences derived from seven clones confirmed the heterozygosity of G081, as they corresponded to two open reading frames of 801 nucleotides that matched either DRB40103101 or the recently described DRB401033, for which a partial sequence, spanning exons 2 and 3, has been reported. The phenotype of G081 (A01; B0702, 1302/1303; Cw0602, 07; DRB10403, 0701; DRB40103101, 01033; DQB10202, 0302; DQA10201, 0301) is consistent with a proposed association of DRB401033 with DRB10403 and DQB1*0302.