We have used a combination of chromosome-mediated gene transfer and microcell fusion techniques to transfer Chinese hamster chromosome 1 to mouse cells. Microcell hybrids containing a single hamster chromosome were analyzed to map genes on this chromosome. We have confirmed the assignment of seven markers (GSR, NP, EST-D, ADK, PEP-S, PGM2, and PEP-B) to hamster chromosome 1. Segregation among the linked markers was induced by X irradiation followed by selection for the retention or loss of human hprt. Cosegregation of markers in independent subclones made it possible to determine the gene order for the seven loci. The gene order proposed for these loci is as follows: pter-GSR-NP-EST-D-ADK-(PEP-S, PGM2)-PEP-B-qter. In addition GSR, NP, EST-D, and ADK have been assigned to pter-1q12; PEP-S and PGM2 to 1q12-1q21, and PEP-B to 1q32-1qter. These regional assignments and gene order on chromosome 1 have provided the information relevant to the linkages conserved between Chinese hamster, mouse, and man.