Chapter 17 Zebrafish/Mouse Somatic Cell Hybrids for the Characterization of the Zebrafish Genome

Publisher Summary This chapter overviews the techniques pertinent to the production and the characterization of zebrafish/mouse cell hybrids. Transfer of zebrafish chromosomes to cells of a foreign species has been accomplished successfully with three different fish cell lines-ZF4, LFF, and AB9—as donor cells, and with the mouse B78 cells or hamster Wg3H cells as recipients. The mouse B78 melanoma cell line was chosen as recipient in whole-cell fusion experiments because it was previously shown to fuse very easily with other cell lines. Furthermore, the cells have a fast growth rate and are easy to maintain, characteristics that were often retained after fusion. Zebrafish chromosomes are tagged with the neo resistance gene. Mouse B78 cells are not deficient in any enzyme—therefore, the aminoglycoside phosphotransferase (neo) gene, conferring resistance to the aminoglycoside drug G418, is introduced into zebrafish chromosomes to select against unfused recipient mouse cells and to ensure retention of zebrafish chromosomes in hybrids.