Comparison of mutant frequencies at the transgenic lambda LacI and cII/cI loci in control and ENU-treated Big Blue mice.
We compared the lambda cII/cI transgenic mutation assay described by Jakubczak et al. [(1996): Proc Natl Acad Sci USA 93:9073-9078] to the previously established Big Blue assay. Genomic DNA isolated from liver, spleen, and lung tissue of control or ethylnitrosourea (ENU)-treated Big Blue mice (100 mg/kg i.p., single dose) was packaged into phage (five animals, two packagings per DNA sample) which were simultaneously plated for lacI and cII/cI mutant frequency (MF) and titer. Mean MF of control animals was higher for cII/cI than lacI for all three tissues examined (spontaneous cII/cI MF divided by spontaneous lacI MF = 2.9, 3.1, and 1.7 for liver, spleen, and lung, respectively). The differences were statistically significant for liver and spleen, but not lung. The ENU-induced MF measured by subtracting control MFs from ENU-treated MFs was higher in the cII/cI assay than lacI (liver = 23.0 x 10(-5) for cII/cI vs. 15.1 x 10(-5) for lacI; spleen = 64.8 x 10(-5) for cII/cI vs. 36.1 x 10(-5) for lacI; lung = 17.1 x 10(-5) for cII/cI vs. 15.8 x 10(-5) for lacI). Fold increase over control values measured by dividing MF of ENU-treated animals by appropriate control values was higher for lacI than cII/cI (liver = 4.4-fold for lacI vs. 2.7 for cII/cI; spleen = 13.1-fold for lacI vs. 8.4 for cII/cI; and lung = 5.6-fold for lacI vs. 4.0 for cII/cI). Despite these differences, overall results were similar for the two mutational endpoints. These results suggest that the cII/cI assay may be an acceptable alternative to lacI where transgenic mutation studies are indicated.
Zimmer
D M
DM
Investigative Toxicology Unit, Pharmacia & Upjohn, Kalamazoo, Michigan 49001, USA.
Harbach
P R
PR
Mattes
W B
WB
Aaron
C S
CS
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D010582:Bacteriophage lambda / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D005038:Ethylnitrosourea / Q000494:pharmacology*; D005819:Genetic Markers / Q000235:genetics; D056968:Lac Repressors; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016296:Mutagenesis / Q000187:drug effects*; D009152:Mutagenicity Tests / Q000379:methods; D012097:Repressor Proteins / Q000235:genetics; D013154:Spleen / Q000187:drug effects / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D019076:Transgenes / Q000235:genetics*; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; D019065:Virus Assembly
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D005819:Genetic Markers; D056968:Lac Repressors; D012097:Repressor Proteins; D014157:Transcription Factors; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C039000:cII protein, bacteriophage lambda; C016649:phage repressor proteins; D005038:Ethylnitrosourea