Stable plastid transformation in PEG-treated protoplasts of Nicotiana tabacum

Plastid engineering currently relies on DNA delivery by the biolistic process. We report here stable plastid transformation in tobacco by an alternate direct transformation protocol that is based on polyethylene glycol (PEG) treatment of leaf protoplasts in the presence of the transforming DNA. Clon...

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Bibliographic Details
Published in:Bio/Technology 1993-01, Vol.11 (1), p.95-97
Main Authors: Golds, T. (Ludwig-Maximilians-University of Munich, Munich, F.R.G.), Maliga, P, Koop, H.U
Format: Article
Language:English
Subjects:
Agriculture
ALCOHOLES
ALCOHOLS
ALCOOL
ANTIBIOTICOS
ANTIBIOTICS
ANTIBIOTIQUE
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
CHEMICAL RESISTANCE
CHLOROPLASTE
CHLOROPLASTS
CLOROPLASTO
DIRECT DNA UPTAKE
DRUGS
Eukaryotic cell cultures
FEUILLE
Fundamental and applied biological sciences. Psychology
GENETIC TRANSFORMATION
GENETICA
GENETICS
GENETIQUE
GENOMAS
GENOME
GENOMES
HOJAS
LEAVES
Life Sciences
MEDICAMENT
MEDICAMENTOS
Methods. Procedures. Technologies
Miscellaneous
NICOTIANA TABACUM
Plant cells and fungal cells
PLASMIDE
PLASMIDIOS
PLASMIDS
PLASTE
PLASTIDIOS
PLASTIDS
POLIETILENO
POLYETHYLENE
POLYETHYLENE GLYCOL
PROTOPLASTE
PROTOPLASTOS
PROTOPLASTS
RESISTANCE AUX PRODUITS CHIMIQUES
RESISTENCIA QUIMICA
SPECTINOMYCIN
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
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Description
Summary:Plastid engineering currently relies on DNA delivery by the biolistic process. We report here stable plastid transformation in tobacco by an alternate direct transformation protocol that is based on polyethylene glycol (PEG) treatment of leaf protoplasts in the presence of the transforming DNA. Clones with transformed plastid genomes were selected by spectinomycin resistance encoded by a mutant 16S ribosomal RNA gene. Incorporation of the transforming DNA into the plastid genome was confirmed by two unselected markers, streptomycin resistance and a novel PstI site that flank the spectinomycin resistance mutation in plasmid pZS148. Our simple and inexpensive protocol eliminates the dependence on the particle gun for chloroplast transformation and should facilitate applications of plastome engineering in crops.
ISSN:0733-222X
1087-0156
2331-3684
1546-1696
DOI:10.1038/nbt0193-95