Molecular and General Genetics MGG

, Volume 199, Issue 2, pp 169–177 | Cite as

Molecular and general genetics of a hybrid foreign gene introduced into tobacco by direct gene transfer

  • Ingo Potrykus
  • Jerzy Paszkowski
  • Michael W. Saul
  • Jirina Petruska
  • Raymond D. Shillito
Article

Summary

Two clones of N. tabacum, transformed to kanamycin resistance by direct transfer to protoplasts of a hybrid gene, consisting of the protein coding region from the bacterial gene for aminoglycoside phosphotransferase under the control of 5′/3′ expression signals from cauliflower mosaic virus gene VI, in the bacterial plasmid pUC8, have been subjected to a detailed genetic crossing analysis accompanied by Southern blot analysis and enzyme activity assays of representative offspring. The genetic data obtained from large populations of R1/F1 and R2/F2 offspring as well as from more than 20 subclones of each of the original transformants confirm that (a) one functional copy of the hybrid gene was stably integrated into chromosomal DNA of the original transformants, (b) that the gene normally was stably maintained during clonal proliferation, (c) that normally it is transmitted in a regular fashion (with exceptions) to sexual offspring, and (d) that it is inherited as a single dominant trait. Data from DNA hybridisation and enzyme assays confirm this interpretation. The functional gene is integrated together with several non-functional copies and bacterial plasmid sequences, which are inherited as one block together with the functional gene.

Keywords

Aminoglycoside Enzyme Activity Assay Cauliflower Mosaic Virus Protein Code Region Hybrid Gene 
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. Chaleff RS (1981) Genetics of higher plants — Application of cell culture. Cambridge University Press, Cambridge London New York New Rochelle Melbourne SydneyGoogle Scholar
  2. De Block M, Herrera-Estrella L, Van Montagu M, Schell J, Zambryski P (1984) Expression of foreign genes in regenerated plants and their progeny. EMBO J 3:1691–1689Google Scholar
  3. Hain R, Stabel P, Czernilofsky AP, Steinbiss H-H, Herrera-Estrella L, Schell J (1985) The use of chimeric genes for the genetic transformation of plant protoplasts by naked DNA. Mol Gen Genet, in pressGoogle Scholar
  4. Horsch RB, Fraley RT, Rogers SG, Sanders PR, Lloyd A, Hoffmann N (1984) Inheritance of functional foreign genes in plants. Science 223:496–498Google Scholar
  5. Lörz H, Baker B, Schell (1985) gene transfer to cereal cells mediated by protoplast transformation Mol Gen Genet 199:178–182Google Scholar
  6. Maliga P, Breznovitz A, Marton L (1973) Streptomycin resistant plants from callus culture of resistant tobacco. Nature 244:29–30Google Scholar
  7. Nitsch JP, Nitsch C (1969) Haploid plants from pollen grains. Science 163:85–87Google Scholar
  8. Paszkowski J, Saul M (1985) Direct gene transfer to plants. Methods Enzymol (in press)Google 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. Perucho M, Hanhan D, Wigler M (1980) Genetic and physical linkage of exogenous sequences in transformed cells. Cell 22:309–317Google Scholar
  11. Potrykus I, Saul MW, Petruska J, Paszkowski J, Shillito RD (1985) Direct gene transfer to cells of a graminaceous monocot. Mol Gen Genet 199:183–188Google Scholar
  12. Reiss B, Sprengel R, Willi M, Schaller H (1984) A new, sensitive method for qualitative and quantitative assay of the neomycin phosphotransferase in crude cell extracts. Gene 30:211–218Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Ingo Potrykus
    • 1
  • Jerzy Paszkowski
    • 1
  • Michael W. Saul
    • 1
  • Jirina Petruska
    • 1
  • Raymond D. Shillito
    • 1
  1. 1.Friedrich Miescher InstitutBaselSwitzerland

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