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  • 1
    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
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  • 2
    ISSN: 1617-4623
    Keywords: Ds1 excision ; Maize Streak Virus ; Zea mays plants ; Agroinfection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A reverse genetic system for studying excision of the transposable elementDs1 in maize plants has been established previously. In this system, theDs1 element, as part of the genome of maize streak virus (MSV), is introduced into maize plants via agroinfection. In the presence of theAc element, excision ofDs1 from the MSV genome results in the appearance of viral symptoms on the maize plants. Here, we used this system to study DNA sequences requiredin cis for excision ofDs1. TheDs1 element contains theAc transposase binding motif AAACGG in only one of its subterminal regions (defined here as the 5′ subterminal region). We showed that mutation of these motifs abolished completely the excision capacity ofDs1. This is the first direct demonstration that the transposase binding motifs are essential for excision. Mutagenesis with oligonucleotide insertions in the other (3′) subterminal region resulted in elements with either a reduced or an increased excision efficiency, indicating that this subterminal region also has an important function.
    Type of Medium: Electronic Resource
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