Abstract
Abstract Mitochondrial DNA can be isotopically labeled to the virtual exclusion of nuclear DNA labeling in cell lines lacking the major soluble thymidine kinase (EC 2.7.1.21) but retaining a mitochondrial thymidine kinase. This characteristic provides a means for the selective assay of mitochondrial DNA during isolation and a determination of the cellular content of mitochondrial DNA. The simplest of the three isolation procedures compared in this study is shown to yield approximately two-thirds of the total mitochondrial DNA in the tissue culture cells examined. Mouse L cells containing predominantly a 107 dalton closed circular mitochondrial DNA species have 1100 ± 250 molecules per cell. A second line of mouse L cells which has mitochondrial DNA of molecular weight 2 x 107 contains approximately 900 molecules per cell. HeLa cells have at least four times the mitochondrial DNA mass per mitochondrial volume as L cells.
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