Satellite DNA in large marker chromosomes of methotrexate-resistant mouse cells

The additional segments of five large marker chromosomes of methotrexate-resistant mouse melanoma PG19T3 cells are shown to consist of C-banding material. In situ hybridization indicates that these additional segments also contain a high proportion of sequences that will cross hybridize with mouse satellite DNA. Analytical density gradient centrifugation suggests that up to 60% of the DNA in the additional segments may be in the form of satellite DNA. Incorporation of bromodeoxyuridine for one complete S phase and staining with the Hoechst 33258 fluorescence plus Giemsa technique reveals complex asymmetries within the additional segments. These asymmetries are interpreted as showing a large repeating unit, which is likely to be involved in the amplification of the dihydrofolate reductase gene.