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Bacillus Subtilis Rec-Assay With and Without Metabolic Activation: Improvements and Applications

  • Tsuneo Kada
  • Susumu Watanabe

Abstract

Chemicals that give positive effects in the rec-assay are often mutagens, chromosome aberrants, and/or carcinogens. Characteristics of the Bacillus subtilis rec-assay system are the following:
  1. 1.

    This assay is one of the systems that is most inexpensive, rapid, and nonlaborious.

     
  2. 2.

    Bacillus subtilis, gram-positive, has a permeable cell membrane to most chemicals. A rec deficiency, rec45, has been selected among DNA repair-deficient markers because of wide spectra of sensitivities to many chemical mutagens.

     
  3. 3.

    Recent improvements using spores in place of vegetative cells enabled us to increase assay sensitivity (some 50-fold), to use S-9 (rat liver homogenate) for in vitro metabolic activation as well as to obtain reproducible quantitative data.

     
The rec-assay can be used very efficiently in combination with current bacterial mutation assays such as the Ames Salmonella system. Some examples of screening of environmental mutagens will be shown as to metal compounds, pesticides, mycotoxins, and food additives.

Keywords

Bacillus Subtilis Inhibition Zone Sodium Nitrite Sorbic Acid Chemical Mutagen 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Ames, B. N., F. D. Lee, and W. E. Durston, 1973. An improved bacterial test system for the detection and classification of mutagens and carcinogens. Proc. Natl. Acad. Sci. (USA), 70: 782.ADSCrossRefGoogle Scholar
  2. Hayatsu, H., K. C. Chung, T. Kada, and T. Nakajima, 1975. Generation of mutagenic compound(s) by a reaction between sorbic acid and nitrite. Mutation Res., 30: 417.CrossRefGoogle Scholar
  3. Hirano, K., K. Narui, Y. Sadaie, and T. Kada, 1978. Some improvements in the mutagenicity testing of chemicals with Bacillus subtilis, Salmonella typhimurium and Escherichia coli; results with 500 chemicals. Mutation Res., 53: 200.Google Scholar
  4. Kada, T., K. Hirano, and Y. Shirasu, 1980. Screening of environmental chemical mutagens by the rec-assay system with Bacillus subtilis, pp. 149-173. In: “Chemical Mutagens,” Vol. 6, “Principles and Methods for their Detection,” F. J. de Serres and A. Hollaender (eds.). Plenum Press, New York.CrossRefGoogle Scholar
  5. Kada, T., K. Morita, and R. Inoue, 1978. Anti-mutagenic action of vegetable factor(s) on the mutagenic principle of tryptophan pyrolysate. Mutation Res., 53: 351.Google Scholar
  6. Kada, T., K. Tutikawa, and Y. Sadaie, 1972. In vitro and host mediated “rec-assay” procedures for screening chemical mutagens; and phloxine, a mutagenic red dye detected. Mutation Res., 16: 165.CrossRefGoogle Scholar
  7. Kada, T., M. Moriya, and Y. Shirasu, 1974. Screening of pesticides for DNA interactions by “rec-assay” and mutagenesis testing, and frameshift mutagens detected. Mutation Res., 26: 243.CrossRefGoogle Scholar
  8. Kada, T., 1973a. DNA-damaging products from reaction between sodium nitrite and sorbic acid. Ann. Rep. Natl. Inst. Genet., 24: 43.Google Scholar
  9. Kada, T., 1973b. Escherichia coli mutagenicity of furylfuramide. Jap. J. Genetics, 48: 301.CrossRefGoogle Scholar
  10. Kada, T., 1976. Rec-assay with cold incubation with and without metabolic activation in vitro. Mutation Res., 38: 340.Google Scholar
  11. Kanematsu, N., M. Hara, and T. Kada, 1980. Rec-assay and mutagen icity studies on metal compounds. Mutation Res., 11 (2): 109–116.Google Scholar
  12. Kondo, S., H. Ichikawa, K. Iwo, and T. Kato, 1970. Base-change mutagenesis and prophage induction in strains of Escherichia coli with different DNA repair capacities. Genetics, 66: 187.Google Scholar
  13. Morita, K., M. Hara, and T. Kada, 1978. Studies on natural des- mutagens: screening for vegetable and fruit factors active in inactivation of mutagenic pyrolysis products from amino acids. Agric. Biol. Chem., 42: 1235.CrossRefGoogle Scholar
  14. Namiki, M. and T. Kada, 1975. Formation of ethylnitolic acid by the reaction of sorbic acid with sodium nitrite. Agric. Biol. Chem., 39: 1335.CrossRefGoogle Scholar
  15. Nishioka, H., 1975. Mutagenicity of metal compounds. Mutation Res., 31: 185.Google Scholar
  16. Sadaie, Y., and T. Kada, 1976. Recombination-deficient mutants of Bacillus subtilis. J. Bacteriol., 125: 489.Google Scholar
  17. Shirasu, Y. M. Moriya, K. Kato, and T. Kada, 1976. Mutagenicity screening of pesticides in the microbial system. Mutation Res., 40: 19–30.CrossRefGoogle Scholar
  18. Shirasu, Y., M. Moriya, K. Kato, F. Lienard, H. Tezuka, S. Teramoto and T. Kada, 1977. Mutagenicity screening of pesticides and modification productsHHHBasis of carcinogenicity evaluation. In: “Origin of Human Cancer,” H. H. Hiatt, J. D. Watson, and J. A. Winsten (eds.). Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.Google Scholar
  19. Sugimura, T., M. Nagao, T. Kawachi, M. Honda, T. Yahagi, Y. Seino, S. Sato, N. Matsukura, T. Matsushima, and A. Shirai, 1977. Mutagenic carcinogens in food with special reference to highly mutagenic pyrolytic products in broiled foods, pp. 1561–1577. In: “Origin of Human Cancer,” H. H. Hiatt, J. D. Watson, and J. A. Winsten (eds.). Cold Spring Harbor Laboratory Symposium, Cold Spring Harbor, New York.Google Scholar
  20. Suzuki, K. and T. Mitsuoka, 1978. Mutagenicity of intestinal bacteria. Mutation Res., 50: 295.CrossRefGoogle Scholar
  21. Ueno, Y., and K. Kubota, 1976. DNA-attacking ability of carcinogenic mycotoxins in recombination-deficient mutant cells of Bacillus subtilis. Cancer Res., 36: 445.Google Scholar

Copyright information

© Plenum Press, New York 1983

Authors and Affiliations

  • Tsuneo Kada
    • 1
  • Susumu Watanabe
    • 1
  1. 1.Department of Induced MutationNational Institute of GeneticsMisima Sizuoka-kenJapan

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