Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1964 May;87(5):1118–1122. doi: 10.1128/jb.87.5.1118-1122.1964

USE OF THYMINELESS DEATH TO ENRICH FOR DOUBLY AUXOTROPHIC MUTANTS OF BACILLUS MEGATERIUM

J T Wachsman a,1, L Hogg a
PMCID: PMC277155  PMID: 4959796

Abstract

Wachsman, J. T. (University of Illinois, Urbana), and L. Hogg. Use of thymineless death to enrich for doubly auxotrophic mutants of Bacillus megaterium. J. Bacteriol. 87:1118–1122. 1964.—When strain KM:T−, a thymine auxotroph of Bacillus megaterium strain KM, is allowed to undergo thymineless death on a minimal medium, the survivors are greatly enriched in polyauxotrophic mutants. Cells were irradiated with ultraviolet light, grown in the presence of thymidine and a complete amino acid mixture, and then starved for thymidine in the absence of amino acids. Doubly auxotrophic mutants (thymine− amino acid−) may account for more than 90% of the survivors. The most reproducible results were obtained when sucrose (0.4 m) was added to both growth and starvation media. Although the percentage of mutants among the survivors increases with the time of thymine starvation, the absolute number of double auxotrophs per milliliter decreases. It is probable that the extent of cross-feeding determines both the mutant yield and the mutants types. Substrains of KM:T− having additional requirements for each of the following amino acids have been isolated: histidine, threonine, tyrosine, tryptophan, arginine, isoleucine, methionine, serine, and cysteine.

Full text

PDF
1118

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ADELBERG E. A., COUGHLIN C. A. Bacterial mutation induced by thymine starvation. Nature. 1956 Sep 8;178(4532):531–532. doi: 10.1038/178531a0. [DOI] [PubMed] [Google Scholar]
  2. BARNER H. D., COHEN S. S. The induction of thymine synthesis by T2 infection of a thymine requiring mutant of Escherichia coli. J Bacteriol. 1954 Jul;68(1):80–88. doi: 10.1128/jb.68.1.80-88.1954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BARNER H. D., COHEN S. S. The isolation and properties of amino acid requiring mutants of a thymineless bacterium. J Bacteriol. 1957 Sep;74(3):350–355. doi: 10.1128/jb.74.3.350-355.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. BAUMAN N., DAVIS B. D. Selection of auxotrophic bacterial mutants through diaminopimelic acid or thymine deprival. Science. 1957 Jul 26;126(3265):170–170. doi: 10.1126/science.126.3265.170. [DOI] [PubMed] [Google Scholar]
  5. GORINI L., KAUFMAN H. Selecting bacterial mutants by the penicillin method. Science. 1960 Feb 26;131(3400):604–605. doi: 10.1126/science.131.3400.604. [DOI] [PubMed] [Google Scholar]
  6. KANAZIR D. The apparent mutagenicity of thymine deficiency. Biochim Biophys Acta. 1958 Oct;30(1):20–23. doi: 10.1016/0006-3002(58)90235-x. [DOI] [PubMed] [Google Scholar]
  7. KELLENBERGER E., LARK K. G., BOLLE A. Amino acid dependent control of DNA synthesis in bacteria and vegetative phage. Proc Natl Acad Sci U S A. 1962 Oct 15;48:1860–1868. doi: 10.1073/pnas.48.10.1860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LATHAM A. B., WEINBERG R. Apparent mutagenic effect of thymine deficiency for a thymine-requiring strain of Escherichia coli. J Bacteriol. 1956 Oct;72(4):570–572. doi: 10.1128/jb.72.4.570-572.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LUBIN M. Enrichment of auxotrophic mutant populations by recycling. J Bacteriol. 1962 Mar;83:696–697. doi: 10.1128/jb.83.3.696-697.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LUBIN M. Selection of auxotrophic bacterial mutants by tritium-labeled thymidine. Science. 1959 Mar 27;129(3352):838–839. doi: 10.1126/science.129.3352.838. [DOI] [PubMed] [Google Scholar]
  11. MAALOE O., HANAWALT P. C. Thymine deficiency and the normal DNA replication cycle. I. J Mol Biol. 1961 Apr;3:144–155. doi: 10.1016/s0022-2836(61)80041-7. [DOI] [PubMed] [Google Scholar]
  12. MANGALO R., WACHSMAN J. T. Effect of 8-azaguanine on growth and viability of Bacillus megaterium. J Bacteriol. 1962 Jan;83:27–34. doi: 10.1128/jb.83.1.27-34.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. WACHSMAN J. T., MANGALO R. Use of 8-azaguanine for the isolation of auxotrophic mutants of Bacillus megaterium. J Bacteriol. 1962 Jan;83:35–37. doi: 10.1128/jb.83.1.35-37.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wachsman J. T., Kemp S., Kogg L. Thymineless death in Bacillus megaterium. J Bacteriol. 1964 May;87(5):1079–1086. doi: 10.1128/jb.87.5.1079-1086.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES