TitleReversion-reporter transgenes to analyze all six base-substitution pathways in Arabidopsis.
Publication TypeJournal Article
Year of Publication2011
AuthorsBollmann SR, Tominey CM, Hoffman PD, Hoffman TMC, Hays JB
JournalPlant Physiol
Date Published2011 Mar
Alleles, Amino Acid Substitution, Arabidopsis, Base Sequence, Gene Dosage, Genes, Reporter, Genetic Engineering, Glucuronidase, Immunohistochemistry, Ions, Metals, Molecular Sequence Data, Mutagenesis, Mutant Proteins, Mutation, Plants, Genetically Modified, Sequence Analysis, DNA, Transgenes, Ultraviolet Rays

To expand the repertoire of Arabidopsis (Arabidopsis thaliana) mutation-reporter transgenes, we constructed six mutant alleles in the same codon of the β-glucuronidase-encoding GUS transgene. Each allele reverts to GUS+ only via a particular one of the six transition/transversion pathways. AcV5 epitope tags, fused carboxyl terminal to the inactive GUS- proteins, enabled semiquantitative immunoassays in plant protein extracts. Spontaneous G:C→T:A transversions, previously not measured using reporter transgenes, were quite frequent. This may reflect mispairing of adenine with 8-oxoguanine in DNA attacked by endogenous oxyradicals. Spontaneous G:C→A:T was modest and other reversions were relatively low, as reported previously. Frequencies of ultraviolet C-induced TT→TC and TC→TT reversions were both high. With increased transgene copy number, spontaneous G:C→T:A reversions increased but ultraviolet C-induced reversions decreased. Frequencies of some reversion events were reduced among T4 versus T3 generation plants. Based on these and other analyses of sources of experimental variation, we propose guidelines for the employment of these lines to study genotoxic stress in planta.

Alternate JournalPlant Physiol.
PubMed ID21212302
PubMed Central IDPMC3046586
Grant ListT32 ES007060 / ES / NIEHS NIH HHS / United States