Transformation of Mitochondrially Coded Genes into Mammalian Cells Using Intact Mitochondria and Mitochondrial DNA
The antibiotics chloramphenicol (CAP) and efrapeptin (EF) inhibit mitochondrial protein synthesis and ATPase, respectively, and thus kill sensitive (wild-type) mammalian cells. However, a variety of mutant cell lines have been isolated and characterized which are resistant to these otherwise lethal antibiotics. We have utilized two mutant murine cell lines, the AMT-BU-Al (Malech and Wivel, 1976) and TL-1 (Lichtor and Getz, 1978), to investigate the possibility of isolating mitochondria from the resistant cells and using them to confer antibiotic resistance on sensitive cells. In addition, experiments were conducted comparing the ability of intact mitochondria or cloned mitochondrial DNA to confer antibiotic resistance on sensitive cells. It is expected that these techniques will be of use not only to those studying mammalian mitochondrial genetics and the mechanisms of DNA-mediated gene transfer, but also to those involved in elucidating the mechanisms by which cytoplasts exert effects on nuclear-coded functions.
KeywordsAntibiotic Resistance Recipient Cell Mycoplasma Contamination Chloramphenicol Resistance Intact Mitochondrion
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