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  • 1
    Electronic Resource
    Electronic Resource
    Transformation of Aspergillus awamori by Agrobacterium tumefaciens –mediated homologous recombination (1999)
    [s.l.] : Nature America Inc.
    Nature biotechnology 17 (1999), S. 598-601 
    Details
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Agrobacterium tumefaciens is known to transfer part of its tumor-inducing (Ti) plasmid to the filamentous fungus Aspergillus awamori by illegitimate recombination with the fungal genome. Here, we show that when this Ti DNA shares homology with the A. awamori genome, integration can also occur by ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/9915
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  • 2
    Electronic Resource
    Electronic Resource
    Agrobacterium tumefaciens -mediated transformation of filamentous fungi (1998)
    [s.l.] : Nature Publishing Company
    Nature biotechnology 16 (1998), S. 839-842 
    Details
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Agrobacterium tumefaciens transfers part of its Ti plasmid, the T-DNA, to plant cells during tumorigenesis. It is routinely used for the genetic modification of a wide range of plant species. We report that A. tumefaciens can also transfer its T-DNA efficiently to the filamentous fungus ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/nbt0998-839
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  • 3
    Electronic Resource
    Electronic Resource
    Cre/lox-mediated recombination in Arabidopsis: evidence for transmission of a translocation and a deletion event (2000)
    Springer
    Chromosoma 109 (2000), S. 287-297 
    Details
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Cre recombinase was used to mediate recombination between a chromosomally introduced loxP sequence in Arabidopsis thaliana (35S-lox-cre) and transferred DNA (T-DNA) originating from Agrobacterium tumefaciens (plox-npt), carrying a single loxP sequence. Constructs were designed for specific Cre-mediated recombination between the two lox sites, resulting in restoration of neomycin phosphotransferase (nptII) expression at the target locus. Kanamycin resistant (Kmr) recombinants were obtained with an efficiency of about 1% compared with random integration. Molecular analyses confirmed that these were indeed due to recombination between the lox sites of the target and introduced T-DNA. However, polymerase chain reaction analysis revealed that these reflected site-specific integration events only in a minority (4%). The other events were classified as translocations/inversions (71%) or deletions (25%), and were probably caused by site-specific recombination between a randomly integrated T-DNA and the original target locus. We studied some of these events in detail, including a Cre-mediated balanced translocation event, which was characterized by a combination of molecular, genetic and cytogenetic experiments (fluorescence in situ hybridization to spread pollen mother cells at meiotic prophase I). Our data clearly demonstrate that Agrobacterium-mediated transfer of a targeting T-DNA with a single lox site allows the isolation of multiple chromosomal rearrangements, including translocation and deletion events. Given that the complete sequence of the Arabidopsis genome will have been determined shortly this method has significant potential for applications in functional genomics.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004120000079
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  • 4
    Electronic Resource
    Electronic Resource
    Improvements in the transformation of Arabidopsis thaliana C24 leaf-discs by Agrobacterium tumefaciens (1996)
    Springer
    Plant cell reports 15 (1996), S. 572-577 
    Details
    ISSN: 1432-203X
    Keywords: Abbreviations:2,4-D: 2,4-dichlorophenoxyacetic acid, AGM:Arabidopsis growth medium, BAP: benzylaminopurine, CaMV: Cauliflower Mosaic Virus, CTAB: Hexadecyltrimethylammonium-bromide, DIG: digoxigenin, FeNaEDTA: Iron-sodium-ethylene-dinitrilo tetraacetic acid complex, GUS:β-Glucuronidase, IBA: indole-3-butyric acid, LB: left T-DNA border, MES: 2-(N-morpholino) ethane sulfonic acid, MS: Murashige and Skoog medium, NAA:α-naphthaleneacetic acid, RB: right T-DNA border
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary. We report here an efficient Arabidopsis leaf-disc transformation protocol yielding an average transformation frequency of 1.6 transgenic shoots per leaf explant 4 weeks after the bacterial infection period. Subsequent cultivation in vitro is such that a high percentage (85–90%) of the primary transformants produces seeds with an average seed yield of 100–300 seeds per plant. This improved transformation protocol yields mainly (70%) transformants segregating for a single T-DNA locus of which 68% actually contain one T-DNA insert. The objective is to generate a pool of independent transformants harboring an activator T-DNA construct in a gene tagging approach to isolate genes involved in morphogenesis and auxin signal transduction.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050076
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  • 5
    Electronic Resource
    Electronic Resource
    Selection of Arabidopsis mutants overexpressing genes driven by the promoter of an auxin-inducible glutathione S-transferase gene (1999)
    Springer
    Plant molecular biology 39 (1999), S. 979-990 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; glutathione S-transferase ; auxin-responsive gene expression ; developmental mutants ; GST ; gup mutant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic arabidopsis plants were isolated that contained a T-DNA construct in which the promoter of an auxin-inducible glutathione S-transferase (GST) gene from tobacco was fused to the kanamycin resistance (nptII) as well as to the β-glucuronidase (gusA) reporter gene. Subsequently, seeds were treated with EMS to obtain mutants in which both reporter gene fusions were up-regulated. Northern analysis showed that the mRNA level of a related, endogenous auxin-inducible GST gene of Arabidopsis was increased in some of these mutants as well. Two of the gup (GST up-regulated) mutants were characterized in more detail and roughly mapped. Both had epinastic cotyledons and leaves, a phenotype that turned out to be linked to the gup mutation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006129426712
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  • 6
    Electronic Resource
    Electronic Resource
    Isolation and characterization of cDNA clones corresponding with mRNAs that accumulate during auxin-induced lateral root formation (1999)
    Springer
    Plant molecular biology 39 (1999), S. 273-287 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; auxin-induced ; glycine-rich protein ; lateral root ; proline-rich protein ; subtilisin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Lateral root formation in root cultures of Arabidopsis thaliana can be initiated by exogenous addition of auxin. In order to find cDNA clones of which the corresponding mRNAs accumulate during this process, a cDNA library was constructed from root cultures treated with the active auxin 1-naphthaleneacetic acid (1- NAA). Differential screening of this library with cDNA probes derived from mRNA populations isolated from root cultures treated with 1-NAA and the inactive analogue 2-naphthaleneacetic acid (2-NAA) led to the isolation of four cDNA clones, designated AIR1, AIR3, AIR9 and AIR12. Accumulation of the mRNAs starts between 4 and 8 h and continues till at least 24 h after addition of an active auxin. Sequence analysis revealed that AIR1 encodes a protein that is related to a large family of proteins that consist of a proline-rich or glycine- rich N-terminus and a hydrophobic, possibly membrane spanning C- terminus. The putative function of these proteins is coupling of the cell wall to the plasma membrane. Surprisingly, AIR1 lacks the proline-rich or glycine-rich N-terminus which is thought to be important for interaction with the cell wall. AIR3 encodes a subtilisin-like serine protease which is believed to be active outside the plant cell. Although AIR9 and AIR12 do not show any significant homology to sequences in the database, they are also predicted to function outside the cell. Our screening thus indicates that a variety of genes encoding extracellular proteins are activated during auxin-induced lateral root formation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006104205959
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  • 7
    Electronic Resource
    Electronic Resource
    Cre/lox-mediated site-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana by transient expression of cre (1998)
    Springer
    Plant molecular biology 38 (1998), S. 393-406 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium ; Arabidopsis ; Cre/lox ; recombination ; site-specific integration ; T-DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Cre/lox system was used to obtain targeted integration of an Agrobacterium T-DNA at a lox site in the genome of Arabidopsis thaliana. Site-specific recombinants, and not random events, were preferentially selected by activation of a silent lox-neomycin phosphotransferase (nptII) target gene. To analyse the effectiveness of Agrobacterium-mediated transfer we used T-DNA vectors harbouring a single lox sequence (this vector had to circularize at the T-DNA left- and right-border sequences prior to site-specific integration) or two lox sequences (this vector allowed circularization at the lox sequences within the T-DNA either prior to or after random integration, followed by targeting of the circularized vector), respectively. Furthermore, to control the reversibility of the integration reaction, Cre recombinase was provided transiently by using a cotransformation approach. One precise stable integrant was found amongst the recombinant calli obtained after transformation with a double-lox T-DNA vector. The results indicate that Agrobacterium-mediated transformation can be used as a tool to obtain site-specific integration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006024500008
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  • 8
    Electronic Resource
    Electronic Resource
    Cre/lox-mediated site-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana by transient expression of cre (1998)
    Springer
    Plant molecular biology 38 (1998), S. 1269-1269 
    Details
    ISSN: 1573-5028
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005842114047
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