NOVEL HLA-LINKED DNA IN A YEAST ARTIFICIAL CHROMOSOME

ALTHOUGH A LARGE NUMBER OF DNA CLONES EXIST WHICH COVER MANYOF THE GENES OF THE HUMAN MAJOR HISTOCOMPATIBILITY COMPLEX (MHC), A NUMBER OF LARGE GAPS EXIST, INCLUDING THE 275-300 KB REGION OF DNA BETWEEN HUMAN LEUCOCYTE ANTIGEN (HLA)-DR AND THE COMPLEMENT GENES. THE MHC ENCODES GENES DETERMING SUSCEPTIBILITY TO DISEASE, INCLUDING JUVENILE-ONSET DIABETES, GLUTEN-SENSITIVE ENTEROPATHY, AND THE BLISTERING SKIN DISEASE PEMPHIGUS VULGARIS. THESE SUSCEPTIBILITY GENESARE EXPECTED TO INCLUDE NOT ONLY IDENTIFIED GENES (CLASS I AND II HLA, COMPLEMENT FACTORS 2, B, AND 4, TUMOR NECROSIS FACTOR A, AND 21-HYDROXYLASE), BUT OTHER AS-YET-UNIDENTIFIEDGENES IN THIS REGION OF EXTENDED HAPLOTYPE WHICH INCLUDES 2500 KILOBASES OF DNA. THIS PROPOSAL DESCRIBES THE PREPARATION OF A DERIVATIVE OF THE PLASMID PYAC3 CONTAINING UNIQUE MLU I AND SAC II SITES, AND THE CLONING OF OVERLAPPING MLU I- AND SAC II-RESTRICTION FRAGMENTS FROM THE HLA-DR-COMPLEMENT GAP AS YEAST ARTIFICIAL CHROMOSOMES (YAC). WE ENVISAGE POTENTIAL APPLICATIONS OF SUBCLONES FOR HUMAN GENETICS, CLINICAL AND FORENSIC.