Human RNA transcripts in man-mouse somatic cell hybrids. II. Thermal denaturation studies and cot analysis

Abstract

The present report confirms an earlier finding that human RNA transcripts from a man-mouse somatic cell hybrid that has regularly retained only the human X chromosome can be identified by molecular hybridization on nitrocellulose filters. From thermal denaturation studies it is concluded that molecular hybrids between hybrid cell and human nucleic acids have higher T _m's, and thus greater specificity, than those between mouse and human nucleic acids. Additional data, utilizing the kinetics of molecular hybridization in solution (Cot analysis), demonstrate the presence of “few gene copy” DNA sequences in the above-mentioned hybrid cells that are complementary to human (HeLa) HnRNA. Also shown is a fraction of hybrid cell HnRNA that is complementary to “few gene copy” DNA sequences of human DNA. Thus, the experiments reported may represent a specific assay for identifying reiterated and “few gene copy” DNA sequences of the human X chromosome and their complementary RNA. Theoretically this experimental approach may be extended to any chromosome. Some of the short-term and long-term perspectives of these types of studies are discussed.