Cybrid formation with recipient cell lines containing dominant phenotypes.
A clone of Chinese hamster ovary (CHO) cells, BT3, resistant to Tevenel, the sulfamoyl analog of chloramphenicol has been isolated. Resistance was found to be at the mitochondrial level and was shown to be cytoplasmically inherited. This marker was then used to develop a method by which a cell line possessing a dominant nuclear mutation (resistance to 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole, DRB) could be used as the recipient in cybrid formation. The unique feature in this procedure was the removal of nucleated cells from the cytoplasts by passage through unipore filters. The dominant character of the DRB- and Tevenel-resistant phenotypes permitted the selection of cybrids immediately after fusion. This initially increased the frequency of cybrid clones 16-fold as compared to a recipient cell line possessing a recessive marker. The possibility of extending the method to recipient cells lacking a selectable drug-resistance marker is discussed.
D000818:Animals; D002460:Cell Line; D002701:Chloramphenicol / Q000031:analogs & derivatives* / Q000494:pharmacology; D002999:Clone Cells / Q000187:drug effects*; D006224:Cricetinae; D003412:Cricetulus; D003584:Cytological Techniques; D004272:DNA, Mitochondrial / Q000235:genetics*; D004351:Drug Resistance; D005111:Extrachromosomal Inheritance; D005260:Female; D005821:Genetic Techniques; D006822:Hybrid Cells / Q000187:drug effects*; D010053:Ovary; D010641:Phenotype
D004272:DNA, Mitochondrial; C003423:tevenel; D002701:Chloramphenicol