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  • 1
    Electronic Resource
    Electronic Resource
    Visualisation of transgene expression at the single protoplast level (1992)
    Springer
    Plant cell reports 11 (1992), S. 346-350 
    Details
    ISSN: 1432-203X
    Keywords: Beet Necrotic Yellow Vein Virus ; Chenopodium quinoa ; Coat protein ; Frequency of expression ; Nicotiana tabacum ; NPT II ; Protoplast printing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts are currently used to study the expression of genes following transformation. Expression is followed on a population of protoplasts after total protein extraction by conventional western blotting or measure of the enzymatic activity of the transgenic protein. We describe here a new method, called protoplast printing, allowing easy detection of the fraction of cells expressing a certain protein within a population of protoplasts. It consists of immobilization of the protoplast proteins on a nitrocellulose filter, so as to retain the outlines of the cell, followed by immunological detection of the protein of interest. The only special requirement is an antibody specific for the protein. We have studied the expression of the BNYVV coat protein after electroporation of Chenopodium quinoa protoplasts with viral RNAs, and the expression of the NPT II gene in protoplasts isolated from transgenic tobacco plants as well as after direct transfer of plasmid DNA into tobacco protoplasts. In both cases — infection with viral RNAs and transformation with plasmid DNA — expressing and non-expressing cells can be distinguished as early as 12h after transfer of the transgenes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233363
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Expression of foreign genes in sunflower (Helianthus annuus L.) — evaluation of three gene transfer methods (1955)
    Springer
    Euphytica 85 (1955), S. 63-74 
    Details
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; electroporation ; particle gun ; polyethylene glycol ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Suitable sunflower tissues and cells were transformed either by direct gene transfer into protoplasts, particle bombardment, or Agrobacterium co-culture. While all techniques allowed efficient short-term or transient expression of the introduced gene(s) in the respective tissues, stable transformation was only observed after transformation with Agrobacterium. The latter technique was suitable for the production of transgenic callus from seedling cotyledons and occasional shoots with chimaeric expression of the transgene. Detailed analysis of the interaction of Agrobacterium with this explant showed that infection efficiency was critically dependent on the co-culture conditions, and that the preferentially-transformed cells were not the ones competent for regeneration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023931
    Library Location Call Number Volume/Issue/Year Availability
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    Paper (German National Licenses)
    Fulltext
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