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Background mutations and polymorphisms in lacZ-plasmid transgenic mice.

M E Dollé1, W K Snyder, N J van Orsouw

  • 1Institute for Drug Development, Cancer Therapy and Research Center, San Antonio, Texas 78229, USA. mdolle@saci.org

Environmental and Molecular Mutagenesis
|October 26, 1999
PubMed
Summary
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Investigating the lacZ-plasmid transgenic mouse model revealed artifactual mutations. These artifacts, mainly from star activity during plasmid recovery, occur at a frequency of 1.3 x 10(-5).

Area of Science:

  • Genetics
  • Molecular Biology
  • Toxicology

Background:

  • Transgenic animal models with reporter genes are crucial for measuring in vivo mutant frequencies.
  • The lacZ-plasmid model excels at detecting large genomic rearrangements, unlike bacteriophage lambda systems.

Purpose of the Study:

  • To investigate background mutant frequencies and spectra in the lacZ-plasmid transgenic mouse model.
  • To identify and quantify sources of artifactual mutations within this model system.

Main Methods:

  • Analysis of mutant frequencies and spectra in lacZ-plasmid transgenic mice.
  • Investigation of host cell mutations (galE, galK, galT) contributing to background.
  • Assessment of plasmid excision procedures, including HindIII restriction enzyme activity.

Related Experiment Videos

Main Results:

  • A subset of mutants (~1.3 x 10^-5) are likely artifacts, independent of true mutation frequency.
  • Host cell mutations (galactose-insensitive) contribute a small fraction to the background.
  • Star activity during HindIII digestion is the largest contributor to background artifacts (~1.1 x 10^-5).
  • Ten polymorphic sites were identified within the lacZ-plasmid cluster in founder line 60.

Conclusions:

  • The lacZ-plasmid transgenic mouse model is sensitive to large rearrangements but susceptible to artifacts.
  • Star activity during DNA recovery is a significant source of background mutations.
  • Understanding these artifacts is crucial for accurate in vivo mutation frequency assessment.