Transient gene expression in sunflower (Helianthus annuus L.) following microprojectile bombardment

Hunold, Reiner; Burrus, Monique; Bronner, Roberte; Duret, Jean-Pierre; Hahne, Günther

doi:10.1016/0168-9452(94)04038-i

Cited by 28 publications

(13 citation statements)

References 42 publications

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“…For biolistic transfection, BY-2 cells were bombarded as described by Hunold et al (1995) with modifications (Haas et al, 2005) and observed 4 h after bombardment.…”

Section: Transient and Stable Cell Transformationsmentioning

confidence: 99%

The Plant TPX2 Protein Regulates Prospindle Assembly before Nuclear Envelope Breakdown

et al. 2008

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The Targeting Protein for Xklp2 (TPX2) is a central regulator of spindle assembly in vertebrate cells. The absence or excess of TPX2 inhibits spindle formation. We have defined a TPX2 signature motif that is present once in vertebrate sequences but twice in plants. Plant TPX2 is predominantly nuclear during interphase and is actively exported before nuclear envelope breakdown to initiate prospindle assembly. It localizes to the spindle microtubules but not to the interdigitating polar microtubules during anaphase or to the phragmoplast as it is rapidly degraded during telophase. We characterized the Arabidopsis thaliana TPX2-targeting domains and show that the protein is able to rescue microtubule assembly in TPX2-depleted Xenopus laevis egg extracts. Injection of antibodies to TPX2 into living plant cells inhibits the onset of mitosis. These results demonstrate that plant TPX2 already functions before nuclear envelope breakdown. Thus, plants have adapted nuclear-cytoplasmic shuttling of TPX2 to maintain proper spindle assembly without centrosomes.

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“…For biolistic transfection, BY-2 cells were bombarded as described by Hunold et al (1995) with modifications (Haas et al, 2005) and observed 4 h after bombardment.…”

Section: Transient and Stable Cell Transformationsmentioning

confidence: 99%

The Plant TPX2 Protein Regulates Prospindle Assembly before Nuclear Envelope Breakdown

et al. 2008

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“…Cells were subcultured every 7 days and harvested 3 days after medium renewal for biolistic bombardment. The harvested cells were filtered onto Whatman discs and placed on 0.8% agar Murashige-Skoog media plates supplemented with 0.25 M mannitol for 2-4 h. Particle preparation and biolistic assays were performed as described (35) with the following modifications: 4 mg of 1.1-m tungsten particles (Bio-Rad) were sterilized in 1 ml of absolute alcohol for 20 min. Particles were then mixed with 10 g of plasmid DNA supplemented with 18% glycerol, 1.5 M CaCl 2 , and 90 mM spermidine in a final volume of 180 l. The firing distance was 11 cm and helium pressure 7 bars.…”

Section: By-2 Transient Expression and Confocal Laser Scanning Microsmentioning

confidence: 99%

The LIM Domains of WLIM1 Define a New Class of Actin Bundling Modules

Thomas

Moreau

Dieterle

et al. 2007

Journal of Biological Chemistry

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“…Cells were filtered onto Whatman disks and placed for 2 to 4 h on 0.8% agar MS media plates supplemented with 0.1 M mannitol and 0.1 M sorbitol. Particle preparation and bombardment assays were performed as described by Hunold et al (1995) with modifications: 2 mg of 1.1 mm tungsten particles (BioRad, Hercules, CA) were immersed in 1 mL of absolute alcohol for 20 min. Dried particles were then successively mixed with 10 mg of recombinant plasmid DNA (pCK-EGFP vector) supplemented with 18% glycerol, 0.75 M CaCl 2 , and 90 mM spermidine in a final volume of 90 mL.…”

Section: Transient Expression In Tobacco By-2 Cellsmentioning

confidence: 99%

The Open Reading Frame VI Product ofCauliflower mosaic virusIs a Nucleocytoplasmic Protein: Its N Terminus Mediates Its Nuclear Export and Formation of Electron-Dense Viroplasms

et al. 2005

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The Cauliflower mosaic virus (CaMV) open reading frame VI product (P6) is essential for the viral infection cycle. It controls translation reinitiation of the viral polycistronic RNAs and forms cytoplasmic inclusion bodies (viroplasms) where virus replication and assembly occur. In this study, the mechanism involved in viroplasm formation was investigated by in vitro and in vivo experiments. Far protein gel blot assays using a collection of P6 deletion mutants demonstrated that the N-terminal a-helix of P6 mediates interaction between P6 molecules. Transient expression in tobacco (Nicotiana tabacum) BY-2 cells of full-length P6 and P6 mutants fused to enhanced green fluorescent protein revealed that viroplasms are formed at the periphery of the nucleus and that the N-terminal domain of P6 is an important determinant in this process. Finally, this study led to the unexpected finding that P6 is a nucleocytoplasmic shuttle protein and that its nuclear export is mediated by a Leu-rich sequence that is part of the a-helix domain implicated in viroplasm formation. The discovery that P6 can localize to the nucleus opens new prospects for understanding yet unknown roles of this viral protein in the course of the CaMV infection cycle.

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Transient gene expression in sunflower (Helianthus annuus L.) following microprojectile bombardment

Cited by 28 publications

References 42 publications

The Plant TPX2 Protein Regulates Prospindle Assembly before Nuclear Envelope Breakdown

The Plant TPX2 Protein Regulates Prospindle Assembly before Nuclear Envelope Breakdown

The LIM Domains of WLIM1 Define a New Class of Actin Bundling Modules

The Open Reading Frame VI Product ofCauliflower mosaic virusIs a Nucleocytoplasmic Protein: Its N Terminus Mediates Its Nuclear Export and Formation of Electron-Dense Viroplasms

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