Conditional, recombinase‐mediated expression of genes in plant cell cultures

Joubès, Jérôme; Schutter, Kristof De; Verkest, Aurine; Inzé, Dirk; Veylder, Lieven De

doi:10.1111/j.1365-313x.2004.02004.x

Cited by 52 publications

(39 citation statements)

References 44 publications

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“…To overcome this difficulty, we decided to use a switch-on constitutive overexpression approach, which relies on CRE-mediated recombination at lox sites (Joubè s et al, 2004) (see Supplemental Figure 5 online). In this design, CRE recombinase expression is controlled by the promoter of the heat-shock protein gene hsp90 and, hence, can be induced by heat treatment.…”

Section: Induced Expression Of Wee1 Induces a G2 Cell Cycle Arrestmentioning

confidence: 99%

ArabidopsisWEE1 Kinase Controls Cell Cycle Arrest in Response to Activation of the DNA Integrity Checkpoint

Schutter¹,

Joubès²,

et al. 2007

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Upon the incidence of DNA stress, the ataxia telangiectasia-mutated (ATM) and Rad3-related (ATR) signaling kinases activate a transient cell cycle arrest that allows cells to repair DNA before proceeding into mitosis. Although the ATM-ATR pathway is highly conserved over species, the mechanisms by which plant cells stop their cell cycle in response to the loss of genome integrity are unclear. We demonstrate that the cell cycle regulatory WEE1 kinase gene of Arabidopsis thaliana is transcriptionally activated upon the cessation of DNA replication or DNA damage in an ATR-or ATM-dependent manner, respectively. In accordance with a role for WEE1 in DNA stress signaling, WEE1-deficient plants showed no obvious cell division or endoreduplication phenotype when grown under nonstress conditions but were hypersensitive to agents that impair DNA replication. Induced WEE1 expression inhibited plant growth by arresting dividing cells in the G2-phase of the cell cycle. We conclude that the plant WEE1 gene is not rate-limiting for cycle progression under normal growth conditions but is a critical target of the ATR-ATM signaling cascades that inhibit the cell cycle upon activation of the DNA integrity checkpoints, coupling mitosis to DNA repair in cells that suffer DNA damage.

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Section: Induced Expression Of Wee1 Induces a G2 Cell Cycle Arrestmentioning

confidence: 99%

ArabidopsisWEE1 Kinase Controls Cell Cycle Arrest in Response to Activation of the DNA Integrity Checkpoint

Schutter¹,

Joubès²,

et al. 2007

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“…Both gfp and nptII genes were driven by CaMV 35S promoters in plasmids pJCGLOX,0 (Figure 1) (Joubès, 2004).…”

Section: Agrobacterium Strains and Plasmidmentioning

confidence: 99%

“…Transformation efficiency of a specific plant can be evaluated conveniently with the gfp gene as a result of a simple assay using a fluorescence microscope. We therefore utilized a binary vector pJCGLOX,0 (Joubès, 2004) that contains a modified gfp gene (egfp) as a reporter and neomycin phosphotransferase II (nptII) as a selectable marker to develop a transformation protocol for Anthurium andraeanum. The effects of explant sources and AS concentrations on transgenic Anthurium andraeanum yield are also discussed.…”

Section: Introductionmentioning

confidence: 99%

Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter

Zhao¹,

Ji²,

Wang³

et al. 2010

Electron. J. Biotechnol.

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Although Agrobacterium-mediated transformation protocols for many economically important plant species have been well established, protocol for a number of flowering plants including Anthurium andraeanum remains challenging. In this study, we report success in generating transgenic Anthurium andraeanum cv Arizona using Agrobacterium GV3101 strain harboring a binary vector carrying gfp as a reporter gene. The possibility of facilitating the screening process for transgenic plants expressing functional proteins using gfp marker was explored. In order to realize high transformation efficiency, different explant sources including undifferentiated callus pieces and petioles were compared for their regeneration efficiency and susceptibility to Agrobacterium-mediated transformation. We also optimized the concentration of AS added to co-cultivation media. Genomic PCR revealed that 11 of the 22 resistant plantlets regenerated on selective medium were successfully transformed. Green fluorescence was observed using a fluorescence microscope in 7 of the 11 PCR-positive plants, indicating GFP was expressed stably in the transformed Anthurium andraeanum. The highest transformation efficiency obtained in this study was 1.71% (percentage of explants with transgenic shoots in total explants) when callus explants were used as starting material and 125 μmol l -1 AS was added during the co-cultivation process.

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“…The wild-type and mutant promoter fragments were subsequently recombined into the pKGWFS7 vector by attL 3 attR recombination reaction at the GATEWAY recombination site located in front of the gene coding for the enhanced green fluorescent protein-GUS (Karimi et al, 2002), resulting in the pCDKBWT and pCDKBmut vectors, respectively. The vectors were used to transform tobacco (Nicotiana tabacum) BY-2 cells as described (Joubè s et al, 2004). For both constructs, 14 calli were selected whose GUS activity was measured as described (Zambre et al, 2003).…”

Section: Promoter Analysismentioning

confidence: 99%

The Plant-Specific Cyclin-Dependent Kinase CDKB1;1 and Transcription Factor E2Fa-DPa Control the Balance of Mitotically Dividing and Endoreduplicating Cells in Arabidopsis

et al. 2004

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Transgenic Arabidopsis thaliana plants overproducing the E2Fa-DPa transcription factor have two distinct cell-specific phenotypes: some cells divide ectopically and others are stimulated to endocycle. The decision of cells to undergo extra mitotic divisions has been postulated to depend on the presence of a mitosis-inducing factor (MIF). Plants possess a unique class of cyclin-dependent kinases (CDKs; B-type) for which no ortholog is found in other kingdoms. The peak of CDKB1;1 activity around the G2-M boundary suggested that it might be part of the MIF. Plants that overexpressed a dominant negative allele of CDKB1;1 underwent enhanced endoreduplication, demonstrating that CDKB1;1 activity was required to inhibit the endocycle. Moreover, when the mutant CDKB1;1 allele was overexpressed in an E2Fa-DPa–overproducing background, it enhanced the endoreduplication phenotype, whereas the extra mitotic cell divisions normally induced by E2Fa-DPa were repressed. Surprisingly, CDKB1;1 transcription was controlled by the E2F pathway, as shown by its upregulation in E2Fa-DPa–overproducing plants and mutational analysis of the E2F binding site in the CDKB1;1 promoter. These findings illustrate a cross talking mechanism between the G1-S and G2-M transition points

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Conditional, recombinase‐mediated expression of genes in plant cell cultures

Cited by 52 publications

References 44 publications

ArabidopsisWEE1 Kinase Controls Cell Cycle Arrest in Response to Activation of the DNA Integrity Checkpoint

ArabidopsisWEE1 Kinase Controls Cell Cycle Arrest in Response to Activation of the DNA Integrity Checkpoint

Optimization in Agrobacterium-mediated transformation of Anthurium andraeanum using GFP as a reporter

The Plant-Specific Cyclin-Dependent Kinase CDKB1;1 and Transcription Factor E2Fa-DPa Control the Balance of Mitotically Dividing and Endoreduplicating Cells in Arabidopsis

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