Techniques for Isolating Nuclear Hybrids
Nuclear-cytoplasmic interactions have been studied in mammalian cells through the formation of cybrids (cytoplasts fused to whole cells) using a mutation for cytoplasmically inherited chloramphenicol resistance (CAPΓ) (Bunn et al., 1974; Wallace et al., 1975; Wallace and Eisenstadt, 1979). McBurney and Strutt (1979) fused karyoplasts to whole cells, a product they termed karyobrids. However, their isolation technique did not distinguish nuclear hybrids from whole-cell hybrids. Muggleton-Harris and Palumbo (1979) produced nuclear hybrids, identified on the basis of nuclear morphology. Because both of these procedures lacked genetic verification, we have developed genetic and physical-genetic techniques for isolating nuclear hybrids. These techniques permit the isolation and verification of nuclear hybrids with the cytoplasmic and mitochondrial content of predominantely one cell.
KeywordsFusion Product Restriction Enzyme Analysis Minimal Fluorescence Nuclear Donor Cell BioI
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