title: Cloned mouse lymphocytes permit analysis of somatic mutations that occur in vivo.
abstract: As part of our mouse model of somatic mutation, we have begun to characterize spontaneously occurring hypoxanthine phosphoribosyltransferase (HPRT) -deficient mouse lymphocytes. Lymphocytes were cloned by in vitro exposure of spleen cells from male C57B1/6 mice to the mitogen concanavalin A, conditioned medium containing lymphocyte growth factors, and thioguanine (TG), in a limiting dilution assay. The 17 TG-resistant clones recovered were all highly deficient in HPRT activity and were found by analysis of surface antigens to be representative of the major subclasses of T lymphocytes. Southern analysis of lymphocyte genomic DNA detected alterations of the hprt gene in 12/17 of the HPRT-deficient lymphocyte clones. Of these 12, 2/17 were lacking the entire hprt locus, 7/17 lacked part of the locus, and 3/17 had other, unidentified alterations.
authors.affiliation: Biomedical Sciences Division, Lawrence Livermore National Laboratory, California 94550.
authors.forename: I M
authors.initials: IM
authors.lastname: Jones
authors.forename: K
authors.initials: K
authors.lastname: Burkhart-Schultz
authors.forename: T L
authors.initials: TL
authors.lastname: Crippen
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000235:genetics; D002999:Clone Cells; D003470:Culture Media; D004247:DNA / Q000235:genetics*; D004252:DNA Mutational Analysis; D004262:DNA Restriction Enzymes; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008957:Models, Genetic; D009154:Mutation; D051381:Rats; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D013866:Thioguanine / Q000378:metabolism
chemical_list: D000954:Antigens, Surface; D003470:Culture Media; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase; D004262:DNA Restriction Enzymes; D013866:Thioguanine