SummaryLinkage crosses were used to determine the gene order of aminoacylase-1, beta-galactosidase, and transferrin on mouse Chromosome 9. The most likely gene order, which was more than 107times more likely than the next most likely order, was transferrin, aminoacylase-1, beta-galactosidase. These results demonstrate that aminoacylase-1 and beta-galactosidase mark a chromosomal segment that has been conserved since divergence of lineages leading to man and mouse. The length of this segment is about 11 cM and the estimate of the true length of the conserved segment marked by these two genes is 33 cM. A second aminoacylase allozyme, which requires MgCl2, is described.