An Experimental Solution for the Luria-Delbrück Fluctuation Problem in Measuring Hypermutation Rates

A cell line harboring all trans-acting elements necessary for hypermutation was transfected with a plasmid harboring the major cis-acting elements plus a green fluorescent protein gene containing a premature chain-termination codon. Transfected cells do not fluoresce unless the stop codon reverts. W...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1999-06, Vol.96 (12), p.6847-6849
Main Authors: Bachl, Jürgen, Dessing, Mark, Olsson, Carina, Von Borstel, R. C., Steinberg, Charles
Format: Article
Language:English
Subjects:
B lymphocytes
Biological Sciences
Cell Line, Transformed
Cell lines
Cells
Codon, Terminator - genetics
Computer Simulation
Cultured cells
Genes
Genetic Markers
Genetic mutation
Genetics
Green Fluorescent Proteins
Humans
Luminescent Proteins - genetics
Lymphocytes
Models, Genetic
Mother cells
Mutation
Plasmids
Population growth
Probability
Proteins
Statistical discrepancies
Stop codon
Transfection
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Summary:A cell line harboring all trans-acting elements necessary for hypermutation was transfected with a plasmid harboring the major cis-acting elements plus a green fluorescent protein gene containing a premature chain-termination codon. Transfected cells do not fluoresce unless the stop codon reverts. When a sizable cell population is purged of revertants by sorting, the frequency of mutants increases linearly with time, and there is no Luria-Delbrück fluctuation effect. Moreover, as mutant frequencies seemed to vary less than cell numbers in replicate cultures, it is suggested that hypermutation might not be coupled closely to cell division.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.96.12.6847