High efficiency transient and stable transformation by optimized DNA microinjection into Nicotiana tabacum protoplasts

Kost B, Galli A, Potrykus I, Neuhaus G (1995)


Publication Status: Published

Publication Type: Journal article

Publication year: 1995

Journal

Publisher: Oxford University Press (OUP): Policy B - Oxford Open Option B / Oxford University Press (OUP)

Book Volume: 46

Pages Range: 1157-1167

Journal Issue: 290

DOI: 10.1093/jxb/46.9.1157

Abstract

Protoplasts were immobilized above a grid pattern and the location of injected cells was recorded by Polaroid photography, The fate of particular targeted cells could be observed, isolation and individual culture of clones derived from injected cells was possible. Following cytoplasmic coinjection of FITC-dextran and 1 mg ml(-1) plasmid DNA on average about 20% of the targeted cells developed into microcalli and roughly 50% of these calli were stably transformed. Transient. expression of the firefly luciferase gene (Luc) was nondestructively analysed 24 h after injection of pAMLuc. Approximately 50% of the injected cells that were alive at this time point expressed the Luc gene transiently. Apparently, stable integration of the injected genes occurred in essentially all transiently expressing cells that developed into clones.

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APA:

Kost, B., Galli, A., Potrykus, I., & Neuhaus, G. (1995). High efficiency transient and stable transformation by optimized DNA microinjection into Nicotiana tabacum protoplasts. Journal of Experimental Botany, 46(290), 1157-1167. https://dx.doi.org/10.1093/jxb/46.9.1157

MLA:

Kost, Benedikt, et al. "High efficiency transient and stable transformation by optimized DNA microinjection into Nicotiana tabacum protoplasts." Journal of Experimental Botany 46.290 (1995): 1157-1167.

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