Detection and analysis of somatic mutations at a lacZ reporter locus in higher organisms: Application to Mus musculus and Drosophila melanogaster

Ana Maria Garcia, Rita A. Busuttil, Armando Rodriguez, Carlos Cabrera, Martha Lundell, Martijn E.T. Dolle, Jan Vijg

Research output: Chapter in Book/Report/Conference proceedingChapter

23 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationBiological Aging
Subtitle of host publicationMethods and Protocols
EditorsTrygve O. Tollefsbol
Pages267-287
Number of pages21
DOIs
StatePublished - Apr 5 2007

Publication series

NameMethods in Molecular Biology
Volume371
ISSN (Print)1064-3745

Keywords

  • Drosophila melanogaster
  • Genome rearrangements
  • Mus musculus
  • Somatic mutations
  • lacZ-plasmids

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Garcia, A. M., Busuttil, R. A., Rodriguez, A., Cabrera, C., Lundell, M., Dolle, M. E. T., & Vijg, J. (2007). Detection and analysis of somatic mutations at a lacZ reporter locus in higher organisms: Application to Mus musculus and Drosophila melanogaster. In T. O. Tollefsbol (Ed.), Biological Aging: Methods and Protocols (pp. 267-287). (Methods in Molecular Biology; Vol. 371). https://doi.org/10.1385/1-59745-361-7:267