Name: a cell-to-cell macromolecular transport assay in planta utilizing biolistic bombardment
Uploaded: 2010-08-27T17:30:00
Description: Watch this Scientific Journal Video about A Cell-to-cell Macromolecular Transport Assay in Planta Utilizing Biolistic Bombardment at JoVE.com

Unsere Arbeit wird durch Zuschüsse von NIH / NIGMS, NSF, USDA / NIFA und BARD zu VC unterstützt.

Hintergrund Symplastisch Transport von Makromolekülen durch Anlage interzellulären Verbindungen, die Plasmodesmen, ist von Interesse für viele Pflanzen-Pathologen und Biologen. Zum Beispiel sind mehrere virale Proteine ​​bekannt, plasmodesmal Größenausschlusschromatographie Grenzen zu regulieren, damit virale Bewegung 1-3. Auch sind einige endogene Proteine, darunter wichtige Entwicklungs-Regulatoren, angenommen von Zelle zu Zelle bewegen, vermutlich durch Plasmodesmen, nic...

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Der Schlüssel für den Erfolg der symplastisch Transport-Assay ist eine hohe Transformationseffizienz, die Produktion von statistisch signifikant und leicht nachweisbaren Signal-Cluster ermöglicht zu erhalten. Dies kann erreicht werden mit den Blättern von gesunden, robusten Pflanzen geerntet, und die Vorbereitung Goldpartikel durch eine reine und konzentrierte DNA-Präparation beschichtet werden. Mit den Blättern an der gleichen Wachstumsphase ist auch entscheidend für den Test Zuverl?...

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<td>Gold microparticles, 1.0 &mu;m in diameter</td>
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a cell-to-cell macromolecular transport assay in planta utilizing biolistic bombardment

Hier stellen wir ein einfaches und schnelles Protokoll zu erkennen und zu beurteilen, das Ausmaß der Zell-Zell-makromolekularen Transport in planta. In diesem Protokoll wird ein fluoreszenzmarkierten-Protein von Interesse vorübergehend in Pflanzengewebe folgenden biolistischen Auslieferung der kodierenden DNA-Konstrukt zum Ausdruck gebracht. Die intra-und interzelluläre Verteilung der markierte Protein wird dann durch konfokale Mikroskopie analysiert. Wir beschreiben diese Technik im Detail und bietet Schritt-für-Schritt-Protokolle Assay und der Beurteilung von symplastisch Transport von Proteinen in drei Pflanzenarten, Arabidopsis thaliana, Nicotiana benthamiana und N. tabacum (Tabak).

Watch this Scientific Journal Video about A Cell-to-cell Macromolecular Transport Assay in Planta Utilizing Biolistic Bombardment at JoVE.com

A Cell-to-cell Macromolecular Transport Assay in Planta Utilizing Biolistic Bombardment

Makromolekulare Handel zwischen Pflanzenzellen können durch transiente Expression eines Fluoreszenz-markierten Proteins von Interesse und die Analyse ihrer intra-und interzelluläre Verteilung durch konfokale Mikroskopie beurteilt werden.

Ein Cell-to-cell Makromolekulare Transport Assay In Planta Verwendung Biolistic Bombardment

Here, we present a simple and rapid protocol to detect and assess the extent of cell-to-cell macromolecular transport in planta. In this protocol, a ...

Research

JoVE Journal

Biology

A Cell-to-cell Macromolecular Transport Assay in Planta Utilizing Biolistic Bombardment

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