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Molecular and General Genetics MGG

, Volume 199, Issue 2, pp 178–182 | Cite as

Gene transfer to cereal cells mediated by protoplast transformation

  • Horst Lörz
  • Barbara Baker
  • Jeff Schell
Article

Summary

Direct gene transfer to cereal cells was achieved by transformation of protoplasts with naked DNA. Protoplasts isolated from cultured cells of Triticum monococcum were incubated in the presence of polyethylene glycol (PEG) with circular and linear plasmid DNA. The pBR322-derived plasmid, pBL1103-4, contained a selectable chimeric gene comprising the protein coding region of the Tn5 aminogly-coside phosphotransferase type II gene (NPT II), the nopaline synthase promoter (pNOS) and the polyadenylation signal of the octopine synthase gene. Transformed cells were selected in medium containing kanamycin and identified by detection of aminoglycoside phosphotransferase II activity.

Keywords

Gene Transfer Kanamycin Polyethylene Glycol Polyadenylation Signal Protein Code Region 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Dudits D, Hadlaczky G, Levi E, Fejer O, Haydu Z, Lazar G (1977) Somatic hybridization of Daucus carota and D. capillifolius by protoplast fusion. Theor Appl Genet 51:127–132Google Scholar
  2. Fedoroff N, Wessler S, Shure S,(1983) Isolation of the transposable controlling elements Ac and Ds. Cell 35:235–242Google Scholar
  3. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158Google Scholar
  4. Hain R, Stabel P, Czernilofsky AP, Steinbiß HH, Herrera-Estrella L, Schell J (1985) Uptake, integration, expression and genetic transmission of a selectable chimeric gene by plant protoplasts. Mol Gen Genet 199:Google Scholar
  5. Hernalsteens JP, Thia-Toong L, Schell J, von Montagu M (1984) An Agrobacterium-transformed cell culture from the monocot Asparagus officinalis. EMBO J 3:3039–3041Google Scholar
  6. Hooykaas-van Slogteren GMS, Hoykaas PJJ, Schilperoort RA (1984) Expression of Ti plasmid genes in monocotyledonous plants infected with Agrobacterium tumefaciens. Nature 311:763–764Google Scholar
  7. Krens FH, Molendijk L, Wullems GJ, Schiperoort RA (1982) In vitro transformation of plant protoplasts with Ti-plasmid DNA Nature 296:72–74Google Scholar
  8. Ozias-Akins P, Lörz H (1984) Progress and limitations in the culture of cereal protoplasts. Trends Biotechnol 2:119–123Google Scholar
  9. Paszkowski J, Shillito RD, Saul M, Mandak V, Hohn T, Hohn B, Potrykus I (1984) Direct gene transfer to plants. EMBO J 3:2717–2722Google Scholar
  10. Potrykus I, Saul M, Petruska J, Paszkowski J, Shillito RD (1985) Direct gene transfer into protoplasts of a graminaceous monocot. Mol Gen Genet 199:183–188Google Scholar
  11. Reiss B, Sprengel R, Will H, Schaller H (1984) A new sensitive method for qualitative and quantitative analysis of neomycin phosphotransferase in crude cell extracts Gene 30:217–223Google Scholar
  12. Schell J, Herrera-Estrella L, Zambryski P, de Block M, Joos H, Willmitzer K, Eckes P, Rosahl S, van Montagu M (1984) Genetic engineering of plants. In: Schell JS, Starlinger P (eds) The impact of gene transfer techniques in eukaryotic cell biology. Springer, Berlin Heidelberg New York, pp 73–90Google Scholar
  13. Shillito RD, Paszkowski J, Potrykus I (1983) Agarose plating and bead type culture techniques enable and stimulate development of protoplast-derived colonies in a number of plant species. Plant Cell Rep 2:244–247Google Scholar
  14. Schreier P, Seftor EA, Schell J, Bohnert HJ (1985) The use of nuclear sequences to direct the light-regulated synthesis and transport of a foreign protein into plant chloroplasts. EMBO J, in pressGoogle Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Horst Lörz
    • 1
  • Barbara Baker
    • 1
  • Jeff Schell
    • 1
  1. 1.Max-Planck-Institut für ZüchtungsforschungKöln 30Federal Republic of Germany

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