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Chemostat selection of Escherichia coli mutants secreting thymidine, cytosine, uracil, guanine, and thymine

  • Applied Genetics and Regulation
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Abstract

Escherichia coli mutants which secreted thymidine, thymine, uracil, cytosine, and guanine into the culture medium were isolated. The isolation strategy was based on the combination of a sensitive screening method and a mutant-generating system. The screening method made use of a thyA mutant of E. coli. These cells, when spread on the agar surface with the 3β-galactosidase indicator X-gal, will grow into bule colonies if a minute amount of thymidine is supplied to them from a nearby secretor colony. A chemostat was used as a mutant-generating system to select for E. coli mutants that were resistant to inhibitors of the pyrimidine biosynthetic pathway. Although many mutants were selected based on their secretion of thymidine, other kinds of nucleosides and nucleobases, such as cytosine, uracil, guanine, and thymine, were also present in larger quantities. This rational selection strategy should be applicable to other species of micro-organisms for the isolation of better producers of nucleosides. The production of nucleosides and nucleobases by fermentation could then become a possibility.

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References

  • Hammer-Jespersen K (1983) Nucleotide catabolism. In Munch-Petersen A (ed) Metabolism of nucleotides, nucleosides and nucleobases in microorganisms. Academic Press, London, pp 203–258

    Google Scholar 

  • Hegeman GD, Rosenberg SL (1970) The evolution of bacterial enzyme systems. Annu Rev Microbiol 24:429–462

    Article  CAS  Google Scholar 

  • Hickson ID, Atkinson KE, Emmerson PT (1982) Molecular cloning and amplification of the gene for thymidylate synthetase of E. coli. Gene 18:257–260

    Article  CAS  Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., pp 433

    Google Scholar 

  • Møllgaard H, Neuhard J (1983) Biosynthesis of deoxythymidine triphosphate. In: Munch-Petersen A (ed) Metabolism of nucleotides, nucleosides and nucleobases in microorganisms. Academic Press, London, pp 149–201

    Google Scholar 

  • Moser H (1958) The dynamics of bacterial population maintained in the chemostat. Carnegie Institution of Washington Publication 614, Washington, D.C.

  • Neuhard J (1983) Utilization of preformed pyrimidine bases and nucleosides. In: Munch-Peterson A (ed) Metabolism of nucleotides, nucleosides and nucleobases in microorganisms. Academic Press, London. pp 95–148

    Google Scholar 

  • Reichard P, Ehrenberg A (1983) Ribonucleotide reductase — a radical enzyme. Science 221: 514–519

    Article  CAS  Google Scholar 

  • Rubin EM, Wilson GA, Young FE (1980) Expression of thymidylate synthetase activity in Bacillus subtilis upon integration of a cloned gene from Escherichia coli. Gene 10: 227–235

    Article  CAS  Google Scholar 

  • Santi DV, McHenry CS (1972) 5-Fluoro-2′-deoxyuridylate: covalent complex with thymidylate synthetase. Proc Natl Acad Sci USA 69: 1855–1857

    Article  CAS  Google Scholar 

  • Schimke R (1982) Gene amplification. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.

    Google Scholar 

  • Stacey KA, Simon E (1965) Improved method for the isolation of thymine-requiring mutants of Escherichia coli. J Bacteriol 90: 554–555

    CAS  PubMed  PubMed Central  Google Scholar 

  • Timson J (1975) Hydroxyurea. Mutat Res 32: 115–132

    Article  CAS  Google Scholar 

  • Williams JW, Morrison JF, Duggleby RG (1979) Methotrexate, a high-affinity pseudosubstrate of dihydrofolate reductase. Biochemistry 12: 2567–2571

    Article  Google Scholar 

  • Yarchoan R, Klecker RW, Weinhold KJ, et al (1986) Administration of 3′-azido-3′-deoxythymidine, an inhibitor of HTLV-III/LAV replication, to patients with AIDS or AIDS-related complex. Lancet: 575–80

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Tsen, S.D. Chemostat selection of Escherichia coli mutants secreting thymidine, cytosine, uracil, guanine, and thymine. Appl Microbiol Biotechnol 41, 233–238 (1994). https://doi.org/10.1007/BF00186965

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  • DOI: https://doi.org/10.1007/BF00186965

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