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Expression of a foreign gene in callus derived from DNA-treated protoplasts of rice (Oryza sativa L.)

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Summary

Protoplasts isolated from suspension cultures of rice cells were treated with bacterial plasmid DNA carrying a chimaeric gene consisting of the nopaline synthase promoter, the aminoglycoside phosphotransferase II (APH(3′)II) structural gene from bacterial transposon Tn5 and the terminator region from cauliflower mosaic virus DNA. Colonies capable of proliferating in medium containing kanamycin (100 μg/ml) were selected. A transformation frequency of approximately 2% to 3% was recorded in several experiments. The enzyme (APH(3′)II) was also detected in kanamycin-resistant callus, which had survived after repeated selection. There was some variation in the APH(3′)II activity in the transformants which paralleled the copy number of the inserted genes.

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References

  • Beck E, Ludwig G, Auerswald EA, Reiss B, Schaller H (1982) Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene 19:327–336

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Lucleic Aids Res 7:1513–1523

    Google Scholar 

  • Depicker A, De Wilde M, De Vos G, De Vos R, Van Montague M, Schell J (1980) Molecular cloning of overlapping segments of the nopaline Ti-plasmid pTIC58 as a means to restriction endonuclease mapping. Plasmid 3:193–211

    Google Scholar 

  • Deshayes A, Herrera-Estrella L, Caboche M (1985) Liposome-mediated transformation of tobacco mesophyll protoplasts by an Escherichia coli plasmid. EMBO J 4:2731–2737

    Google Scholar 

  • Fromm ME, Taylor LP, Walbot V (1986) Stable transformation of maize after gene transfer by electroporation. Nature 319:791–793

    Google Scholar 

  • Gardner RC, Howarth AJ, Hahn P, Brown-Luedi M, Shepherd RJ, Messing J (1981) The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by M13mp7 shotgun sequencing. Nucleic Acids Res 9:2871–2888

    Google Scholar 

  • Hain R, Stabel P, Czernilosky AP, Steinbib HH, Herrera-Estrella L, Schell J (1985) Uptake, integration, expression and genetic transmission of a selectable chimaeric gene by plant protoplasts. Mol Gen Genet 199:161–168

    Google Scholar 

  • Herrera-Estrella L, De Block M, Messens E, Hernalsteens JP, Van Montagu M, Schell J (1983) Chimeric genes as dominant selectable markers in plant cells. EMBO J 2:987–995

    Google Scholar 

  • Krens FA, Molendijk L, Wullems GJ, Schilperoort RA (1982) In vitro transformation of plant protoplast with Ti-plasmid DNA. Nature 296:72–74

    Google Scholar 

  • Leemans J, Langenakens J, DE Greve II, Deblaere R, Van Montagu M, Schell J (1982) Broad-host-range cloning vectors derived from W-plasmid Sa. Gene 19:361–364

    Google Scholar 

  • Lörz H, Baker B, Schell J (1985) Gene transfer to cereal cells mediated by protoplast transformation. Mol Gen Genet 199:178–182

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Google Scholar 

  • Paszkowski J, Shillito RD, Saul M, Mandak V, Hohn T, Hohn B, Potrykus I (1984) Direct gene transfer to plants. EMBO J. 3:2717–2722

    Google Scholar 

  • Potrykus I, Saul MW, Petruska J, Paszkowski J, Shillito RD (1985) Direct gene transfer to cells of a graminaceous monocot Mol Gen Genet 199:183–188

    Google Scholar 

  • Reiss B, Sprengel R, Will H, Schaller H (1984) A new sensitive for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30:211–218

    Google Scholar 

  • Schreier PH, Seftor EA, Schell J, Bohnert HJ (1985) The use of nuclear-encoded sequences to direct the light-regulated synthesis and transport of a foreign protein into plant chloroplasts. EMBO J 4:25–32

    Google Scholar 

  • Southern E (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Uchimiya H, Ohtani T, Ohgawara T, Harada H, Sugita M, Sigiura (1983) Molecular cloning of tobacco chromosomal and chloroplast DNA segments capable of replication in yeast. Mol Gen Genet 192:1–4

    Google Scholar 

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Communicated by H. Saedler

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Uchimiya, H., Fushimi, T., Hashimoto, H. et al. Expression of a foreign gene in callus derived from DNA-treated protoplasts of rice (Oryza sativa L.). Molec. Gen. Genet. 204, 204–207 (1986). https://doi.org/10.1007/BF00425499

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  • DOI: https://doi.org/10.1007/BF00425499

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