Electroporation of Tobacco Leaf Protoplasts Using Plasmid DNA or Total Genomic DNA View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

1995-08-11

AUTHORS

Jac A. Nickoloff , Patrick Gallois , Keith Lindsey , Renee Malone

ABSTRACT

Direct gene transfer into tobacco leaf protoplasts via electroporation is used in many laboratories for transient-expression studies. Protoplasts in general are a convenient model to study events that occur in planta rapidly. Transient expression provides answers in a matter of days instead of the several months that would be needed if the same experiment was carried out with transgenic plants. Examples include: elicitor regulation of a plant defense gene (1), UV induction of chalcone synthase (2), induction of an Adh1 promoter by low concentrations of oxygen (3), hormone response (4,5), transactivation of a promoter by cotransfection with two plasmids, one encoding a transcription factor and the other one carrying its target promoter in front of a reporter gene (6), homologous recombination (7), and nuclear targeting (8,9). However it must be noted that although in the above examples transient expression is mimicking what can be observed in stable transformants, there are cases reported where there are noticeable differences between the two approaches. For example, the strictly seed-specific gene encoding soybean β-conglycinin is expressed in tobacco mesophyll protoplasts, but the sequences responsible for that expression are different from those controlling expression in stable-transformation experiments (10). The same situation has been suggested for the promoter of the pathogenesis-related 1a protein gene (11). More... »

PAGES

89-108

References to SciGraph publications

  • 1994-01. Opportunities for manipulating the seed protein composition of wheat and barley in order to improve quality in TRANSGENIC RESEARCH
  • 1974-01. Plant protoplast fusion and growth of intergeneric hybrid cells in PLANTA
  • 1987. Gene Targeting in Plants in PLANT MOLECULAR BIOLOGY
  • 1991-08. Nuclear DNA content of some important plant species in PLANT MOLECULAR BIOLOGY REPORTER
  • 1988-09. Regulation of a wheat promoter by abscisic acid in rice protoplasts in NATURE
  • 1980-06. Nutritional requirements of protoplast-derived, haploid tobacco cells grown at low cell densities in liquid medium in PLANTA
  • 1980-05. Isolation of the chicken thymidine kinase gene by plasmid rescue in NATURE
  • 1987-09. Hybrid genes in the analysis of transformation conditions in PLANT MOLECULAR BIOLOGY
  • 1987-09. Transfer of a Ten–Member Genomic Library to Plants Using Agrobacterium tumefaciens in NATURE BIOTECHNOLOGY
  • 1987-12. Gene rescue in plants: A model system for “shotgun” cloning by retransformation in MOLECULAR GENETICS AND GENOMICS
  • 1991-03. Electroporated protoplasts express seed specific gene promoters in PLANT CELL REPORTS
  • 1991-10. A DNA Transformation–Competent Arabidopsis Genomic Library in Agrobacterium in BIO/TECHNOLOGY
  • 1990-07. Temporary inhibition of cell wall synthesis improves the transient expression of the GUS gene in Brassica napus mesophyll protoplasts in PLANT CELL REPORTS
  • 1982-04. Isolation and preliminary characterization of a human transforming gene from T24 bladder carcinoma cells in NATURE
  • 1983-09. A plant DNA minipreparation: Version II in PLANT MOLECULAR BIOLOGY REPORTER
  • Book

    TITLE

    Plant Cell Electroporation And Electrofusion Protocols

    ISBN

    0-89603-328-7

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1385/0-89603-328-7:89

    DOI

    http://dx.doi.org/10.1385/0-89603-328-7:89

    DIMENSIONS

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