A DNA fragment of 219 bp was obtained by polymerase chain reaction (PCR) on human genomic DNA using two oligonucleotide mixtures derived from peptide sequences of human placenta choline acetyltransferase (ChAT) and from partially conserved amino acid sequences of rat, porcine and Drosophila ChAT. Sequence homology with porcine ChAT demonstrated that this fragment is part of the human ChAT gene. This gene was assigned to chromosome 10 by hybridization of the 219 bp DNA probe with DNA from human-hamster somatic cell hybrids, and to region 10q11.2-10qter by PCR experiments.