HLA-DR allogenotyping using exon-specific cDNA probes and application of rapid minigel methods.
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.
D000483:Alleles; D003001:Cloning, Molecular; D004247:DNA; D004587:Electrophoresis, Agar Gel; D005091:Exons; D005838:Genotype; D006681:HLA-D Antigens / Q000235:genetics*; D006684:HLA-DR Antigens / Q000235:genetics*; D007158:Immunologic Techniques; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization
D006681:HLA-D Antigens; D006684:HLA-DR Antigens; D004247:DNA