Abstract
An Escherichia coli strain producing transposase of a repeated sequence of Bordetella pertussis chromosome (RSBP) was constructed. A defective MGE-helper plasmid method, which allowed the determination of transposase functional activity was developed. It was shown that transposase synthesized in E. coli cells ensures transposition of “defective” RSBP into the host chromosome. Overexpression of transposase was shown to markedly decrease the vital activity of E. coli cells under selective cultivation conditions. Reasons for a decrease in viability transposase-producing cells are discussed. Results showing the impact of transposase on replication of recombinant plasmids and E. coli cell division were obtained.
Similar content being viewed by others
References
Mahillon, J. and Chandler, M., Insertion Sequences, Microbiol. Mol. Biol. Rev., 1998, vol. 62, pp. 725–774.
Nechaeva, E.V., Volgina, T.I., Kirillov, M.Yu., et al., Mobile Element RSVR1 of Bordetella pertussis, Mol. Genet. Microbiol. Virusol., 1999, vol. 12, pp. 22–27.
McPheat, W.L., Hanson, J.H., Livey, L., et al., Analysis of Separate Isolates of Bordetella pertussis Repeated DNA Sequences, J. Gen. Microbiol., 1989, vol. 135, pp. 1515–1520.
Parkhill, J., Sebaihia, M., Preston, A., et al., Comparative Analysis of the Genome Sequences of Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica, Nat. Genet., 2003, vol. 35, pp. 32–40.
Nechaeva, E.V., Sivov, I.G., and Karataev, G.I., Transposition and Inheritance of a Bordetella Tn Element in Escherichia coli K-12 Cells, Mol. Genet., Mikrobiol. Virusol., 2000, vol. 1, pp. 20–23.
Kirillov, M.Yu., Shumakov, Yu.L., Nechaeva, E.V., et al., Nucleotide Sequence and Properties of an Inverted Repetitive Element of the Bordetella pertussis Chromosome, Genetika (Moscow), 1993, vol. 29, no. 8, pp. 1267–1276.
Lacey, B.W., Antigenic Modulation of Bordetella pertussis, J. Hyg., 1960, vol. 58, pp. 57–93.
Gershanovich, V.N., Bol’shakova, T.N., and Erlagaeva, R.S., Phosphoenolpyruvate-Dependent Phosphotransferase System and Metabolic Regulation in the Bacterial Cell, Usp. Sovrem. Biol., 1987, vol. 103, pp. 173–188.
Shumakov, Yu.L., Kirillov, M.Yu., Nechaeva, E.V., et al., Nucleotide Sequence and Properties of a Transposon-Like Structure cloned from the Bordetella pertussis Chromosome, Genetika (Moscow), 1993, vol. 29, no. 7, pp. 1061–1069.
Maniatis, T., Fritsch, E.F., and Sambrook, J.E., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, New York: Cold Spring Harbor Lab., 1982.
Miller, J.H., Experiments in Molecular Genetics, Cold Spring Harbor, New York: Cold Spring Harbor Lab., 1972.
Vorontsov, V.V., Sivov, I.G., Karataev, G.I., et al., Identification of the Open Reading Frame Coding for Transposase of the Bordetella pertussis RS Element, Mol. Genet., Microbiol. Virusol., 2004, vol. 1, pp. 16–23.
Laemmli, U.K., Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4, Nature, 1970, vol. 227, pp. 680–685.
novagen pET System Manual, 2002, 10th ed.
Sivov, I.G., Bol’shakova, T.N., and Karataev, G.I., Integration and Intramolecular Transposition of the TnBp3 Transposon of Bordetella pertussis in the Escherichia coli K-12 Cells Mutant for the Hpr Protein of the Phosphoenolpyruvate-Dependent Phosphotransferase System, Rus. J. Genet., 2001, vol. 37, no. 7, pp. 741–747.
Kirillov, M.Y., Shumakov, Y., Nechaeva, E.V., et al., Repeated Sequences Isolated from Bordetella pertussis Induce DNA Rearrangements and Deletions at High Frequency, Gene, 1995, vol. 166, pp. 111–116.
Kirillov, M.Yu., Nechaeva, E.V., and Karataev, G.I., Mobile Genetic Element of Bordetella pertussis Chromosome Stimulates Cointegrate Formation in Escherichia coli Strains, Rus. J. Genet., 1997, vol. 33, no. 12, pp. 1384–1390.
Yigit, H. and Reznikoff, W.S., Escherichia coli DNA Topoisomerase I Copurifies with Tn5 Transposase, and Tn5 Transposase Inhibits Topoisomerase I, J. Bacteriol, 1999, vol. 181, pp. 3185–3192.
Yigit, H. and Reznikoff, W.S., Escherichia coli DNA Topoisomerase I and Suppression of Killing by Tn5 Transposase Overproduction: Topoisomerase I Modulates Tn5 Transposition, J. Bacteriol., 1998, vol. 180, pp. 5866–5874.
Yigit, H. and Reznikoff, W.S., Examination of the Tn5 Transposase Overproduction Phenotype in Escherichia coli and Localization of a Suppressor of Transposase Over-production Killing That Is an Allele of rpoH, J. Bacteriol., 1997, vol. 179, pp. 1704–1713.
Author information
Authors and Affiliations
Additional information
Original Russian Text © V.V. Vorontsov, I.G. Sivov, A.M. Umyarov, L.N. Karataev, 2006, published in Genetika, 2006, Vol. 42, No. 1, pp. 39–48.
Rights and permissions
About this article
Cite this article
Vorontsov, V.V., Sivov, I.G., Umyarov, A.M. et al. Functional activity of transposase of Bordetella pertussis IS element in Escherichia coli cells. Russ J Genet 42, 32–39 (2006). https://doi.org/10.1134/S1022795406010042
Received:
Issue Date:
DOI: https://doi.org/10.1134/S1022795406010042