Abstract
Methods to detect and analyze somatic mutations in higher organisms are critically important in view of their causal role in cancer, heritable diseases, and, possibly, aging. Here, we describe detailed protocols for the use of a mutational reporter system based on lacZ-containing plasmids integrated in the germline of Mus musculus and Drosophila melanogaster. Plasmids containing the bacterial lacZ gene integrated at one or more chromosomal sites can be excised, purified and recovered in suitable Escherichia coli hosts allowing the positive selection of mutant lacZ genes and their further molecular characterization. This system is capable of detecting a broad range of mutational events, varying from small mutations in the lacZ reporter gene to large genome rearrangements with one breakpoint in lacZ and the other breakpoint elsewhere in the genome.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Vijg, J. (2000) Somatic mutations and aging: a re-evaluation. Mutat. Res. 447, 117–135.
Vijg, J. and Dollé, M. E. (2002) Large genome rearrangements as a primary cause of aging. Mech. Ageing Dev. 123, 907–915.
Fukuchi, K., Martin, G. M., and Monnat, R. J., Jr. (1989) Mutator phenotype of Werner syndrome is characterized by extensive deletions. Proc. Natl. Acad. Sci. USA 86, 5893–5897.
Gossen, J. A., de Leeuw, W. J., Tan, H., et al. (1989) Efficient rescue of integrated shuttle vectors from transgenic mice: a model for studying mutations in vivo. Proc. Natl. Acad. Sci. USA 86, 7971–7975.
Boerrigter, M. E., Dolle, M. E., Martus, H. J., Gossen, J. A., and Vijg, J. (1995) Plasmid-based transgenic mouse model for studying in vivo mutations. Nature 377, 657–659.
Szilard, L. (1959) On the nature of the aging process. Proc. Natl. Acad. Sci. USA 45, 30–45.
Dollé, M. E., Giese, H., Hopkins, L., Martus, H. J., Hausdorff, J. M., and Vijg, J. (1997) Rapid accumulation of genome rearrangements in liver but not in brain of old mice. Nat. Genet. 17, 431–434.
Dollé, M. E., Snyder, W. Gossen, J. A., Lohman, P. H., and Vijg, J. (2000) Distinct spectra of somatic mutations accumulated with age in mouse heart and small intestine. Proc. Natl. Acad. Sci. USA 97, 8403–8408.
Dollé, M. E. and Vijg, J. (2002) Genome dynamics in aging mice. Genome Res. 12, 1732–1738.
Dollé, M. E., Martus, H. J., Gossen, J. A., Boerrigter, M. E., and Vijg, J. (1996) Evaluation of a plasmid-based transgenic mouse model for detecting in vivo mutations. Mutagenesis 11, 111–118.
Gossen, J. A., Martus, H. J., Wei, J. Y., and Vijg, J. (1995) Spontaneous and X-rayinduced deletion mutations in a LacZ plasmid-based transgenic mouse model. Mutat. Res. 331, 89–97.
Busuttil, R. A., Rubio, M., Dollé, M. E., Campisi, J., and Vijg, J. (2006) Mutant frequencies and spectra depend on growth state and passage number in cells cultured from transgenic lac-plasmid reporter mice. DNA Repair 5(1), 52–60.
Dollé, M. E., Snyder, W. van Orsouw, N. J., and Vijg, J. (1999) Background mutations and polymorphisms in lacZ-plasmid transgenic mice. Environ. Mol. Mutagen 34, 112–120.
Dolle, M. E., Martus, H. J., Novak, M., van Orsouw, N. J., and Vijg, J. (1999) Characterization of color mutants in lacZ plasmid-based transgenic mice, as detected by positive selection. Mutagenesis 14, 287–293.
Dollé, M. E., Busuttil, R. A., Garcia, A. M., et al. (2006) Increased genomic instability is not a prerequisite for shortened lifespan in DNA repair deficient mice. Mutat. Res. 596, 22–35.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Garcia, A.M. et al. (2007). Detection and Analysis of Somatic Mutations at a lacZ Reporter Locus in Higher Organisms. In: Tollefsbol, T.O. (eds) Biological Aging. Methods in Molecular Biology™, vol 371. Humana Press. https://doi.org/10.1007/978-1-59745-361-5_20
Download citation
DOI: https://doi.org/10.1007/978-1-59745-361-5_20
Publisher Name: Humana Press
Print ISBN: 978-1-58829-658-0
Online ISBN: 978-1-59745-361-5
eBook Packages: Springer Protocols