Abstract
Fragile X syndrome and other trinucleotide diseases are characterized by an elongation of a repeating DNA triplet. The ensemble-averaged lambda exonuclease digestion rate of different substrates, including one with an elongated FMR1 gene containing 120 CGG repeats, was measured using absorption and fluorescence spectroscopy. By use of magnetic tweezers sequence-dependent digestion rates and pausing was measured for individual lambda exonucleases. Within the triplet repeats a lower average and narrower distribution of rates and a higher frequency of pausing was observed.
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Acknowledgments
RC acknowledge the support of a National Research Council Research Associateship Award and the help of Karen Usdin and Daman Kumari of NIDDK who supplied the elongated FMR1 sequence and helped with preparation, replication and isolation of the plasmid, and Keir Neuman for critical feedback. This research was supported in part by the Intramural Research Program of the NINDS, NIH.
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Conroy, R.S., Koretsky, A.P. & Moreland, J. Lambda exonuclease digestion of CGG trinucleotide repeats. Eur Biophys J 39, 337–343 (2010). https://doi.org/10.1007/s00249-009-0502-5
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DOI: https://doi.org/10.1007/s00249-009-0502-5