HLA-DR allogenotyping using exon-specific cDNA probes and application of rapid minigel methods

Abstract

The construction of exon-specific HLA-DR beta cDNA clones and their application as DNA or RNA hybridization probes in DR allogenotyping are described. Using a probe derived from the beta 2, transmembrane, cytoplasmic and 3'-untranslated exons of the DR beta chain gene (pRTV1), it is possible to discriminate between DR alleles using the restriction endonucleases TaqI or BamHI. Micro-scale chromosomal DNA isolation procedures, used in conjunction with Southern blot analysis in minigels, facilitate the rapid identification of DR allogenotypes without significant loss of hybridization signal resolution.