TY - JOUR
T1 - Background mutations and polymorphisms in lacZ-plasmid transgenic mice
AU - Dollé, Martijn E.T.
AU - Snyder, Wendy K.
AU - Van Orsouw, Nathalie J.
AU - Vijg, Jan
N1 - Copyright:
Copyright 2007 Elsevier B.V., All rights reserved.
PY - 1999
Y1 - 1999
N2 - Transgenic animal models harboring chromosomally integrated shuttle vectors with bacterial reporter genes are now widely used to measure in vivo mutant frequencies. The lacZ-plasmid transgenic mouse model has a unique sensitivity to large rearrangements compared to systems using bacteriophage lambda vectors, which mainly detect point mutations and small deletions or insertions. In this study, the background mutant frequencies and spectra in the lacZ-plasmid transgenic mouse model were investigated. While the majority of the recovered lacZ-mutants appeared to have originated in the mouse, a subset of mutants are likely to represent artifacts, and occur with a frequency of about 1.3 x 10-5, irrespective of the total mutant frequency. Galactose-insensitive host cells, due to gale back mutations or galK or gait forward mutations, grow through the positive selection system and cause a small subset of the background. When using HindIII to excise the plasmids from genomic DNA, the largest contribution to the background, (1.1 ± 0.3) x 10-5, appeared to be caused by star activity, i.e., cleavage at nucleotide sequences other than the HindIII restriction enzyme recognition sequence, during the recovery procedure. Finally, a total of 10 polymorphic sites in different copies of the lacZ-plasmid cluster in founder line 60 were discovered.
AB - Transgenic animal models harboring chromosomally integrated shuttle vectors with bacterial reporter genes are now widely used to measure in vivo mutant frequencies. The lacZ-plasmid transgenic mouse model has a unique sensitivity to large rearrangements compared to systems using bacteriophage lambda vectors, which mainly detect point mutations and small deletions or insertions. In this study, the background mutant frequencies and spectra in the lacZ-plasmid transgenic mouse model were investigated. While the majority of the recovered lacZ-mutants appeared to have originated in the mouse, a subset of mutants are likely to represent artifacts, and occur with a frequency of about 1.3 x 10-5, irrespective of the total mutant frequency. Galactose-insensitive host cells, due to gale back mutations or galK or gait forward mutations, grow through the positive selection system and cause a small subset of the background. When using HindIII to excise the plasmids from genomic DNA, the largest contribution to the background, (1.1 ± 0.3) x 10-5, appeared to be caused by star activity, i.e., cleavage at nucleotide sequences other than the HindIII restriction enzyme recognition sequence, during the recovery procedure. Finally, a total of 10 polymorphic sites in different copies of the lacZ-plasmid cluster in founder line 60 were discovered.
KW - HindIII
KW - Plasmid
KW - Polymorphism
KW - Star activity
KW - Transgenic
KW - lacZ
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U2 - 10.1002/(SICI)1098-2280(1999)34:2/3<112::AID-EM9>3.0.CO;2-W
DO - 10.1002/(SICI)1098-2280(1999)34:2/3<112::AID-EM9>3.0.CO;2-W
M3 - Article
C2 - 10529734
AN - SCOPUS:0032725343
SN - 0893-6692
VL - 34
SP - 112
EP - 120
JO - Environmental and Molecular Mutagenesis
JF - Environmental and Molecular Mutagenesis
IS - 2-3
ER -