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Big Blue Rodent Cell Cultures for Mutagenesis Studies

I gene of the Big Blue lambda shuttle vector as the target for mutagenesis. After the cells are treated with a test compound, in vitro packaging rapidly and efficiently recovers the lambda shuttle vector from the genomic DNA of the cell. These cell lines exhibit rescue efficiencies between 5x104 and 1x105 plaque-forming units per g of genomic DNA. Bacterial detection systems screen for mutations in the lacI target gene. The target gene can then be recovered from mutant plaques for sequence analysis.

In addition to short-term in vitro mutagenesis studies, these cell lines are useful for various toxicological applications, studies of the effects of oncogenes and tumor suppressor genes on mutation frequencies and studies of DNA repair.

REFERENCES

  1. Kohler, S.W., et al. (1991) Proc. Natl. Acad. Sci. USA 88: 78597962.

  2. Dycaico, M.J., et al. (1994) Mutat. Res. 307: 461478.

  3. Rogers, B.J., et al. (1995) Mutat. Res. 327: 5766.

  4. Dycaico, M.J., et al. (1996) Carcinogenesis 17: 23472356.

  5. Wyborski, D.L., Malkhosyan, S., Moores, J.C., Dycaico, M.J., and Short, J.M. (1994) Strategies 7: 5556.

  6. Wyborski, D.L., et al. (1995) Mutat. Res. 334: 161165.

  7. Manjanatha, M.G., et al. (1996) Environ. Mol. Mutagen. 27: 44


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