, Volume 69, Issue 5, pp 559–564 | Cite as

Characterization of the mutant recombinant strainPseudomonas putida IPM-36 exhibiting anticipating growth on a medium containing an inducer of thecry3A gene expression

  • N. G. Koretskaya
  • O. E. Svetoch
  • O. I. Loseva
  • A. P. Dobritsa
Experimental Articles


Induction of the expression of the 8-endotoxin gene fromBacillus thuringiensis var.tenebrionis in the recombinant strainPseudomonas putida IPM-36 negatively affected the viability and the growth rate of the culture. In order to optimize the insecticide production by the recombinant strain, mutant clones exhibiting anticipating growth on an inducer-containing medium were selected and studied. These clones differed in such aspects as the localization of mutations (either in plasmid pBTN11, carrying thecry3A gene, or in the chromosome), growth rate, or the level of δ-endotoxin synthesis after induction. Several obtained mutants proved much superior toP. putida IPM-36 in their structural and segregation stability, although they were as efficient as the original strain with respect to the production of the insecticide protein Cry3A.

Key words

δ-endotoxin induction gene expression growth rate genetic stability Pseudomonas putida 


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  1. 1.
    Marston, F.A.O., The Purification of Eukaryotic Proteins Synthesized inEscherichia coli, Biochem. J., 1986, vol. 240, pp. 1–12.PubMedGoogle Scholar
  2. 2.
    Vind, J., Sorensen, M.A., Rasmussen, M.D., and Pedersen, S., Synthesis of Proteins inEscherichia coli Is Limited by the Concentration of Free Ribosomes,J. Mol. Biol, 1993, vol. 231, pp. 678–688.PubMedCrossRefGoogle Scholar
  3. 3.
    Weinreich, M.D., Yigit, H., and Reznikoff, W.S., Overexpression of the Tn5 Transposase inEscherichia coli Results in Filamentation, Aberrant Nucleoid Segregation, and Cell Death: Analysis of Escherichia coli and Transposase Suppressor Mutations,J. Bacteriol., 1994, vol. 176, no. 17, pp. 5494–5504.PubMedGoogle Scholar
  4. 4.
    Dong, H., Nilsson, L., and Kurland, C.G., Gratuitous Overexpression of Genes inEscherichia coli Leads to Growth Inhibition and Ribosome Destruction,J. Bacteriol., 1995, vol. 177, no. 6, pp. 1497–1504.PubMedGoogle Scholar
  5. 5.
    Koretskaya, N.G., Svetoch, O.E., Loseva, O.I., and Dobritsa, A.P., Induced Synthesis of Coleoptera-Specific Insecticidal Protein ofBacillus thuringiensis in Cells ofPseudomonas putida, Biotekhnologiya, 1995, vol. 9/10, pp. 8–13.Google Scholar
  6. 6.
    Worsey, M.J. and Williams, P.A., Metabolism of Toluene and Xylenes byPseudomonas putida (arvilla) mt-2: Evidence for a New Function of the TOL Plasmid,J. Bacteriol., 1975, vol. 124, pp. 7–13.PubMedGoogle Scholar
  7. 7.
    Mermod, N., Ramos, J.L., Lehrbach, P.R., and Timmis, K.N., Vector for Regulated Expression of Cloned Genes in a Wide Range of Gram-Negative Bacteria,J. Bacteriol., 1986, vol. 167, no. 2, pp. 447–454.PubMedGoogle Scholar
  8. 8.
    Koretskaya, N.G., Loseva, O.I., Gerasimov, V.N., Pryamchuk, S.D., Svetoch, O.E., and Dobritsa, A.P., Expression of the S-EndotoxincryIIIA Gene ofBacillus thuringiensis in a RecombinantPseudomonas putida Strain under the Control of thePm Promoter and the Regulator GenexylS, Mikrobiologiya, 1998, vol. 67, no. 3, pp. 349–355.Google Scholar
  9. 9.
    Simon, R., Priefer, U., and Puhler, A., A Broad Host-Range Mobilization System forIn Vivo Genetic Engineering: Transposon Mutagenesis in Gram-Negative Bacteria,Bio/technology, 1983, vol. 1, pp. 784–791.CrossRefGoogle Scholar
  10. 10.
    Maniatis, T., Fritsch, E.F., and Sambrook, J.,Molecular Cloning: A Laboratory Manual, Cold Spring Harbor: Cold Spring Harbor Lab., 1982. Translated under the titleMolekulyarnoe klonirovanie, Moscow: Mir, 1984.Google Scholar
  11. 11.
    Pirt, S.J.,Principles of Microbe and Cell Cultivation, Oxford: Blackwell, 1975. Translated under the titleOsnovy kul’tivirovaniya mikroorganizmov i kletok, Moscow: Mir, 1978.Google Scholar
  12. 12.
    Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J., Protein Measurement with the Folin Phenol Reagent,J. Biol. Chem., 1951, vol. 193, pp. 265–275.PubMedGoogle Scholar

Copyright information

© MAIK “Nauka/Interperiodica” 2000

Authors and Affiliations

  • N. G. Koretskaya
    • 1
  • O. E. Svetoch
    • 1
  • O. I. Loseva
    • 1
  • A. P. Dobritsa
    • 1
  1. 1.State Research Center for Applied MicrobiologyObolensk, Moscow oblastRussia

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