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A modified low copy number binary vector pUN for Agrobacterium-mediated plant transformation

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Biologia Plantarum

Abstract

A modified low copy number plant binary vector pUN has been constructed and successfully used to clone unstable DNA sequences. The vector pUN comprises of low copy number, broad host range RK2 replicon from pBin19 and of multiple cloning site (MCS) and T-DNA region, both from a pBINPLUS-derived pLV06 vector. The absence of the ColE1 replicon in the backbone of the binary vector significantly contributed to stability of hardly clonable DNA sequences and enabled their transfer into the tobacco plants through Agrobacterium-mediated transformation.

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Abbreviations

GUS:

β-glucuronidase

MCS:

multiple cloning site

NPTII, NPTIII:

neomycin phosphotransferase II and III

References

  • Bevan, M.: Binary Agrobacterium vectors for plant transformation.-Nucl. Acids Res. 12: 8711–8721, 1984.

    Article  PubMed  CAS  Google Scholar 

  • Cheng, M., Lowe, B.A., Spencer, M., Ye, X., Amstrong, C.L.: Factors influencing Agrobacterium-mediated transformation of monocotyledonous species.-In Vitro cell. dev. Biol. Plant 40: 31–45, 2004.

    Article  Google Scholar 

  • Frary, A., Hamilton, C.M.: Efficiency and stability of high molecular weight DNA transformation: analysis in tomato.-Transgenic Res. 10: 121–132, 2001.

    Article  PubMed  CAS  Google Scholar 

  • Goss, P.J.E., Peccoud, J.: Analysis of the stabilizing effect of Rom on the genetic network controlling ColE1 plasmid replication.-In: Altman, P., Dunker, A.K., Hunter, L., Klein T.E. (ed.): Pacific Symposium on Biocomputing. Pp. 65–76. World Scientific, Hawaii 1999.

    Google Scholar 

  • Hansen, G., Wright, M.S.: Recent advances in the transformation of plants.-Trends Plant Sci. 4: 226–231, 1999.

    Article  PubMed  Google Scholar 

  • Hellens, R., Mullineaux, P., Klee, H.: A guide to Agrobacterium binary Ti vectors.-Trends Plant Sci. 5: 446–451, 2000.

    Article  PubMed  CAS  Google Scholar 

  • Kahn, M., Kolter, R., Thomas, C., Figurski, D., Meyer, R., Remaut, E., Helinski, D.R.: Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2.-Methods Enzymol. 68: 268–280, 1979.

    Article  PubMed  CAS  Google Scholar 

  • Lerner, C.G., Inouye, M.: Low copy number plasmids for regulated low-level expression of cloned genes in Escherichia coli with blue/white insert screening capability.-Nucl. Acids Res. 18: 4631, 1990.

    Article  PubMed  CAS  Google Scholar 

  • Mead, D., Patterson, M, Godiska, R.: High stability for cloning deleterious genes and genomes.-In: Plant & Animal Genomes XIII Conference, San Diego 2005.

  • Mlynarová, L., Libantová, J., Vrba, L., Nap, J.P.: The promiscuity of heterospecific lox sites increases dramatically in the presence of palindromic DNA.-Gene 296: 129–137, 2002.

    Article  PubMed  Google Scholar 

  • Moravčíková, J., Libantová, J., Nap, J.P., Mlynarová, L.: [The influence of the type of the binary vector and the concentration of Agrobacterium inoculum on the expression of reporter gene in transgenic plants.]-In: Užík, M. (ed.): Nové Poznatky z Genetiky a Šl’achtenia Pol’nohospodárskych Rastlín. Pp. 105–106. Výskumný Ústav Rastlinnej Vý roby, Piešt’any 2003. [In Slovak.]

    Google Scholar 

  • Ordway, J.M., Detloff, P.J.: In vitro synthesis and cloning of long CAG repeats.-BioTechniques 21: 609–610, 1996.

    PubMed  CAS  Google Scholar 

  • Pierson, V.L., Barcak, G.J.: Development of E. coli strains tolerating unstable DNA sequences on Col E1 vectors.-Focus 21: 18–19, 1999.

    Google Scholar 

  • Tao, Q., Zhang, H.B.: Cloning and stable maintenance of DNA fragments over 300 kb in Escherichia coli with conventional plasmid-based vectors.-Nucl. Acids Res. 26: 4901–4909, 1998.

    Article  PubMed  CAS  Google Scholar 

  • Trinh, T., Jessee, J., Bloom, R.F., Hirsch, V.: STBL2TM: an Escherichia coli strain for the stable propagation of retroviral clones and direct repeat sequences.-Focus 16: 78–80, 1994.

    Google Scholar 

  • Uranbey, S., Sevimay, C.S., Kaya, M.D., Ipek, A., Sancak, C., Basalma, D., Er, C., Özcan, S.: Influence of different cocultivation temperatures, periods and media on Agrobacterium tumefaciens-mediated gene transfer.-Biol. Plant. 49: 53–57, 2005.

    Article  Google Scholar 

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Authors and Affiliations

  1. Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, P.O. Box 39A, SK-95007, Nitra, Slovak Republic

    E. Vaculková, J. Moravčíková, I. Matušíková & J. Libantová

  2. Research Institute of Animal Production, Hlohovska 2, SK-94992, Nitra, Slovak Republic

    M. Bauer

Authors
  1. E. Vaculková
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  2. J. Moravčíková
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  3. I. Matušíková
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  4. M. Bauer
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  5. J. Libantová
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Correspondence to J. Libantová.

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Vaculková, E., Moravčíková, J., Matušíková, I. et al. A modified low copy number binary vector pUN for Agrobacterium-mediated plant transformation. Biol Plant 51, 538–540 (2007). https://doi.org/10.1007/s10535-007-0116-7

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  • DOI: https://doi.org/10.1007/s10535-007-0116-7

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