Abstract
An experimental assessment was carried out to determine directly the frequency and types of spontaneous base substitutions that occur in chloroplast DNA. A target site within the chloroplast 16S rRNA gene of the green alga Chlamydomonas reinhardtii was chosen for the assay. Mutations at this site were known to confer spectinomycin resistance and simultaneously result in the loss of an AatII cleavage site. In the experiments reported here, base substitutions at any individual base occurred at a frequency in the range of 0.9–11 per 109 viable cells plated. Four new mutations that confer resistance to spectinomycin were identified at the target site in the Chlamydomonas chloroplast 16S rRNA gene. When the relative rates of transition and transversion mutations were quantified, a bias toward transversions was observed. The prominence of A/T → C/G transversions in the observed mutation spectrum suggests that oxidative damage may be the major cause of base substitution mutations within the chloroplast.
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Acknowledgements
We thank Herberto Cerutti for his suggestion regarding the Aat II assay, and Michele Fluck and Ken Keegstra for helpful discussions. This work was supported by grant MCB 9982600 from the National Science Foundation to B.B.S., and was carried out in compliance with current laws governing genetic experimentation in the USA.
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Communicated by R. Herrmann
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GuhaMajumdar, M., Sears, B.B. Chloroplast DNA base substitutions: an experimental assessment. Mol Genet Genomics 273, 177–183 (2005). https://doi.org/10.1007/s00438-005-1121-1
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DOI: https://doi.org/10.1007/s00438-005-1121-1