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Uptake, integration, expression and genetic transmission of a selectable chimaeris gene by plant protoplasts

(1985) MOLECULAR & GENERAL GENETICS. 199(2). p.161-168
Author
Organization
Abstract
Genetic transformation of Nicotiana tabacum protoplasts was achieved by incubation of protoplasts with a plasmid DNA-calcium phosphate coprecipitate, followed by fusion of the protoplasts in the presence of polyvinyl alcohol and subsequent exposure to high pH. A derivative of the plasmid pBR322 containing a chimaeric gene, consisting of the nopaline synthase promoter, the coding region of the aminoglycoside phosphotransferase gene of Tn5 and the polyadenylation signal region of the octopine synthase gene, was used for these transformation experiments. This chimaeric gene confers resistance of transformed plant cells to kanamycin. This novel transformation procedure reproducibly yielded transformants at frequencies of approximately 0.01%. Aminoglycoside phosphotransferase II activity was detected in both transformed calli and in regenerated plants. DNA from some of the transformed clones was analyzed by Southern blot hybridization. The input DNA appears to be integrated into high molecular weight cellular DNA. Genetic analysis of one of the kanamycin resistant plants shows that the chimaeric gene is transmitted to the progeny as a single dominant trait in a Mendelian fashion. As a comparison the input DNA was also introduced into tobacco protoplasts using Agrobacterium tumefaciens and Ti-plasmid derived gene vectors.

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MLA
Hain, Rüdiger, Priska Stabel, Armin P Czernilofsky, et al. “Uptake, Integration, Expression and Genetic Transmission of a Selectable Chimaeris Gene by Plant Protoplasts.” MOLECULAR & GENERAL GENETICS 199.2 (1985): 161–168. Print.
APA
Hain, R., Stabel, P., Czernilofsky, A. P., Steinbiß, H.-H., Herrera-Estrella, L., & Schell, J. (1985). Uptake, integration, expression and genetic transmission of a selectable chimaeris gene by plant protoplasts. MOLECULAR & GENERAL GENETICS, 199(2), 161–168.
Chicago author-date
Hain, Rüdiger, Priska Stabel, Armin P Czernilofsky, Hans-Henning Steinbiß, Luis Herrera-Estrella, and Jeff Schell. 1985. “Uptake, Integration, Expression and Genetic Transmission of a Selectable Chimaeris Gene by Plant Protoplasts.” Molecular & General Genetics 199 (2): 161–168.
Chicago author-date (all authors)
Hain, Rüdiger, Priska Stabel, Armin P Czernilofsky, Hans-Henning Steinbiß, Luis Herrera-Estrella, and Jeff Schell. 1985. “Uptake, Integration, Expression and Genetic Transmission of a Selectable Chimaeris Gene by Plant Protoplasts.” Molecular & General Genetics 199 (2): 161–168.
Vancouver
1.
Hain R, Stabel P, Czernilofsky AP, Steinbiß H-H, Herrera-Estrella L, Schell J. Uptake, integration, expression and genetic transmission of a selectable chimaeris gene by plant protoplasts. MOLECULAR & GENERAL GENETICS. 1985;199(2):161–8.
IEEE
[1]
R. Hain, P. Stabel, A. P. Czernilofsky, H.-H. Steinbiß, L. Herrera-Estrella, and J. Schell, “Uptake, integration, expression and genetic transmission of a selectable chimaeris gene by plant protoplasts,” MOLECULAR & GENERAL GENETICS, vol. 199, no. 2, pp. 161–168, 1985.
@article{6974879,
  abstract     = {Genetic transformation of Nicotiana tabacum protoplasts was achieved by incubation of protoplasts with a plasmid DNA-calcium phosphate coprecipitate, followed by fusion of the protoplasts in the presence of polyvinyl alcohol and subsequent exposure to high pH. A derivative of the plasmid pBR322 containing a chimaeric gene, consisting of the nopaline synthase promoter, the coding region of the aminoglycoside phosphotransferase gene of Tn5 and the polyadenylation signal region of the octopine synthase gene, was used for these transformation experiments. This chimaeric gene confers resistance of transformed plant cells to kanamycin. This novel transformation procedure reproducibly yielded transformants at frequencies of approximately 0.01%. Aminoglycoside phosphotransferase II activity was detected in both transformed calli and in regenerated plants. DNA from some of the transformed clones was analyzed by Southern blot hybridization. The input DNA appears to be integrated into high molecular weight cellular DNA. Genetic analysis of one of the kanamycin resistant plants shows that the chimaeric gene is transmitted to the progeny as a single dominant trait in a Mendelian fashion. As a comparison the input DNA was also introduced into tobacco protoplasts using Agrobacterium tumefaciens and Ti-plasmid derived gene vectors.},
  author       = {Hain, Rüdiger and Stabel, Priska and Czernilofsky, Armin P and Steinbiß, Hans-Henning and Herrera-Estrella, Luis and Schell, Jeff},
  issn         = {0026-8925},
  journal      = {MOLECULAR & GENERAL GENETICS},
  language     = {eng},
  number       = {2},
  pages        = {161--168},
  title        = {Uptake, integration, expression and genetic transmission of a selectable chimaeris gene by plant protoplasts},
  url          = {http://dx.doi.org/10.1007/BF00330254},
  volume       = {199},
  year         = {1985},
}

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