title: Specificity of mutations induced in transfected DNA by mammalian cells.
abstract: DNA transfected into mammalian cells is subject to the high mutation frequency of approximately 1% per gene. We present data bearing on the derivation of the two main classes of mutations detected, base substitutions and deletions. The DNA sequence change is reported for nearly 100 independent base substitution mutations that occurred in shuttle vectors as a result of passage in simian cells. All of the mutations occur at G:C base pairs and involve either transition to A:T or transversion to T:A. To identify possible mutational intermediates, various topological forms of the vector DNA were introduced separately. Supercoiled and relaxed DNA are mutated at equal frequencies. However, linearized DNA leads to a greatly elevated frequency of deletions. Nicked and gapped templates stimulate both deletions and base substitutions. We discuss a model involving intracellular degradation of the transfected DNA which explains these observations.
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002872:Chromosome Deletion; D004247:DNA / Q000235:genetics*; D005822:Genetic Vectors; D008322:Mammals; D008957:Models, Genetic; D009154:Mutation; D010957:Plasmids; D014162:Transfection
chemical_list: D004247:DNA