Somatic cell genetics is an exciting and rapidly expanding field of research. Since descriptions of the major experimental techniques in the field are scattered throughout various journals and other publications, there is a real need for a single reference source for both established investigators and students in the field. In addition, technical reports are frequently abridged such that many researchers are discouraged from attempting to adopt the appropriate methodology. This book, therefore, describes in detail the many recent technical advances in such areas of somatic cell genetics as transfer mediated by liposomes, erythrocyte ghosts, chromosomes, micro cells, mito chondria, and isolated nuclear DNA. These techniques have increased our understanding of the organization and regulation of eukaryotic cells. The production of antibiotic-resistant cell lines and their use in studying cytoplasmic inheritance are also included. Evidence for the cytoplasmic regulation of nuclear gene expression in eukaryotic cells is rapidly accumu lating following the characterization of cytoplasmic mutations. The produc tion of nuclear-coded mutations, their use in standard cell hybridization, and recent advances in techniques for fusing whole cells or cell components are also described. Front Matter....Pages i-xxvi Selection of Purine and Pyrimidine Nucleoside Analog Resistance in Mammalian Cells....Pages 1-9 Techniques for Using HAT Selection in Somatic Cell Genetics....Pages 11-21 Techniques for Decreasing the Toxicity of Polyethylene Glycol....Pages 23-34 The Use of Dimethyl Sulfoxide in Mammalian Cell Fusion....Pages 35-45 The Selection of Heterokaryons and Cell Hybrids Using the Biochemical Inhibitors Iodoacetamide and Diethylpyrocarbonate....Pages 47-65 Techniques for Enucleation of Mammalian Cells....Pages 67-79 Nonselective Isolation of Fibroblast Heterokaryons, Hybrids, and Cybrids by Flow Sorting....Pages 81-100 Techniques for Monitoring Cell Fusion: The Synthesis and Use of Fluorescent Vital Probes (R18, F18)....Pages 101-109 Inheritance of Oligomycin Resistance in Tissue Culture Cells....Pages 111-121 Cytoplasmic Inheritance of Rutamycin Resistance in Mammalian Cells....Pages 123-137 Erythromycin Resistance in Human Cells....Pages 139-157 Cytoplasmic Inheritance of Chloramphenicol Resistance in Mammalian Cells....Pages 159-187 The Influence of Cytoplast-to-Cell Ratio on Cybrid Formation....Pages 189-201 Transformation of Mitochondrially Coded Genes into Mammalian Cells Using Intact Mitochondria and Mitochondrial DNA....Pages 203-210 Mitochondrial Influences in Hybrid Cells....Pages 211-219 Shedding of Tumor Cell Surface Membranes....Pages 221-235 Production of Microcytospheres....Pages 237-243 Isolation and Characterization of Mitoplasts....Pages 245-254 Nuclear Transplantation with Mammalian Cells....Pages 255-267 Techniques for Purifying L-Cell Karyoplasts with Minimal Amounts of Cytoplasm....Pages 269-280 Techniques for Isolating Nuclear Hybrids....Pages 281-289 Monolayer Enucleation of Colcemid-Treated Human Cells and Polyethylene Glycol 400-Mediated Fusion of Microkaryoplasts (Microcells) with Whole Cells....Pages 291-307 Microcell-Mediated Chromosome Transfer....Pages 309-327 Techniques for Isolating Chromosome-Containing Minisegregant Cells....Pages 329-347 Techniques to Isolate Specific Human Metaphase Chromosomes....Pages 349-373 Techniques of Chromosome-Mediated Gene Transfer....Pages 375-383 Transfer of Macromolecules Using Erythrocyte Ghosts....Pages 385-398 Liposome-Mediated DNA Transfer....Pages 399-413 Techniques of DNA-Mediated Gene Transfer for Eukaryotic Cells....Pages 415-428 Viral-DNA Vectors in the Analysis of Mammalian Differentiation....Pages 429-447 Detection of Specific DNA Sequences in Somatic Cell Hybrids and DNA Transfectants....Pages 449-461 Microinjection Turns a Tissue Culture Cell into a Test Tube....Pages 463-470 Techniques of Somatic Cell Hybridization by Fusion of Protoplasts....Pages 471-492 Techniques for Chromosome Analysis....Pages 493-511 Genetic Analysis of Hybrid Cells Using Isozyme Markers as Monitors of Chromosome Segregation....Pages 513-524 Future Perspectives in Somatic Cell Genetics....Pages 525-529 Back Matter....Pages 531-541
