PEG-mediated plastid transformation: a new system for transient gene expression assays in chloroplasts View Full Text


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Article Info

DATE

1991-10

AUTHORS

B. Spörlein, M. Streubel, G. Dahlfeld, P. Westhoff, H. U. Koop

ABSTRACT

Evidence is presented for the introduction of functional copies of the GUS-reporter gene with plastid regulatory signals into chloroplasts after treatment of Nicotiana plumbaginifolia leaf protoplasts with PEG. GUS-activity is found in cells derived from protoplasts treated with PEG in the presence of plasmids harbouring the GUS-gene under the control of plastid promoter and terminator signals (plastid-specific reporter gene constructions). The activity is maintained after chloroplast isolation and incubation with the protease thermolysin under conditions sufficient to completely remove the much higher transient nuclear/cytoplasmic expression of a GUS-gene carrying the CaMV 35S-promoter. Likewise, GUS-activity derived from a plasmid coding for the nuclear/cytoplasmic expression of the reporter gene with a plastid transit presequence is also maintained after these procedures. These results indicate that PEG-treatment is a suitable protocol by which to introduce DNA into chloroplasts for the study of transient gene expression. More... »

PAGES

717-722

References to SciGraph publications

  • 1985-01. Deleted forms of plastid DNA in albino plants from cereal anther culture in CURRENT GENETICS
  • 1987. Plastid Transformation: A Progress Report in PLANT DNA INFECTIOUS AGENTS
  • 1982-11. Lincomycin resistance, a new type of maternally inherited mutation in Nicotiana plumbaginifolia in CURRENT GENETICS
  • 1990-04. Streptomycin and lincomycin resistances are selective plastid markers in cultured Nicotiana cells in MOLECULAR GENETICS AND GENOMICS
  • 1989-02. A cytokinin-sensitive mutant of the moss,Physcomitrella patens, defective in chloroplast division in PROTOPLASMA
  • 1989-05. The amyloplast-targeting transit peptide of the waxy protein of maize also mediates protein transport in vitro into chloroplasts in MOLECULAR GENETICS AND GENOMICS
  • 1985-09. Transcription of the gene encoding the 51 kd chlorophyll a-apoprotein of the photosystem II reaction centre from spinach in MOLECULAR GENETICS AND GENOMICS
  • 1991-07. Transfer of defined numbers of chloroplasts into albino protoplasts using an improved subprotoplast/protoplast microfusion procedure: Transfer of only two chloroplasts leads to variegated progeny in MOLECULAR GENETICS AND GENOMICS
  • 1991-02. Subcellular location and expression level of a chimeric protein consisting of the maize waxy transit peptide and the β-glucuronidase of Escherichia coli in transgenic potato plants in MOLECULAR GENETICS AND GENOMICS
  • 1987-05. High-velocity microprojectiles for delivering nucleic acids into living cells in NATURE
  • 1989-08. PEG- and electroporation-induced transformation in Nicotiana tabacum: influence of genotype on transformation frequencies in THEORETICAL AND APPLIED GENETICS
  • 1984-09. Nucleotide sequence of the clustered genes for apocytochrome b6 and subunit 4 of the cytochrome b/f complex in the spinach plastid chromosome in CURRENT GENETICS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/bf00227316

    DOI

    http://dx.doi.org/10.1007/bf00227316

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1015053158

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/24213446


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