Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica

Cytogenetics and Cell Genetics
T B NesterovaS Zakian

Abstract

Two hundred thirty-seven independent somatic cell hybrids have been obtained between opossum (Monodelphis domestica) splenocytes, bone marrow cells, or primary fibroblasts, and HPRT-deficient or TK-deficient Chinese hamster, mouse, American mink, or common vole fibroblast lines. Because extreme segregation and fragmentation of marsupial chromosomes commonly occurs in eutherian x marsupial somatic cells hybrids, we developed a rapid primary screening method that enables the identification of primary clones containing a large amount of opossum DNA 20-25 d after fusion. This method, which depends on in situ hybridization of biotin-labeled total opossum DNA on interphase nuclei of hybrid cells fixed on the bottom of microwell plates, was used to screen the 237 hybrid clones; 52 of them had a substantial amount of opossum DNA. G-banding and in situ hybridization of biotin-labeled total opossum DNA on metaphase spreads of the clones enabled identification of 17 hybrid clones containing from two to seven intact chromosomes of M. domestica on the background of Chinese hamster or vole chromosomes. The hybrid clones with intact opossum chromosomes are used in a panel constructed for mapping the opossum genome. Initial mapping results fro...Continue Reading

Citations

Mar 3, 2007·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Alexander I ShevchenkoSuren M Zakian
Aug 31, 2001·ILAR Journal·Paul B. Samollow
Aug 31, 2001·ILAR Journal·Jennifer A. Marshall Graves

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