Genomic clones covering the entire sequence of the gene encoding human 2-oxoglutarate dehydrogenase (OGDH) were isolated by screening leukocyte and placenta genomic libraries with radio- and digoxigenin (DIG)-labeled human OGDH cDNAs. The human OGDH gene contains 22 exons spanning approx. 85 kb. All exon/intron splice junctions follow the GT/AG rule. The results of primer extension analysis imply that the OGDH transcription start point is a thymine residue 55 bp upstream from the ATG start codon. The 5'-flanking region of OGDH lacked canonical TATA or CAAT boxes. Using DNAs from human x rodent somatic cell hybrids that segregate human chromosomes in conjunction with fluorescence in situ hybridization, human OGDH was assigned to chromosome arm 7p, an area that corresponds to the boundary between bands 7p13 and 7p14.