Humana Press eBooks · 2006 · 109 citations · 21 references
Life SciencesPlant BiologyPlant SciencesGene ExpressionPlant BiotechnologyPlant GeneticsAgrobacterium-mediated TransformationPlant EnzymesPlant Gene ExpressionPlant TranscriptomicsReporter Gene AssaysReverse Transcription Polymerase Chain Reaction (Rt-pcr)Transgene ExpressionTransgene Expression AnalysisTransient ExpressionTransient Expression SystemsTransient Gene ExpressionWestern Blot Analysis
An Agrobacterium-mediated transient expression assay has been described for in vivo analysis of constitutive or inducible gene expression in Arabidopsis plants. By simple infiltration of Agrobacterium cells carrying appropriate gene constructs into Arabidopsis leaves, transient expression assays can be performed within 3 d without using expensive instruments or complicated procedures. Two days after agroinfiltration, Arabidopsis plants can be treated with chemical inducers such as dexamethasone to induce transgene expression. The expression of the beta-glucoronidase (GUS) reporter gene can be determined by histochemical staining. In addition, RNA and protein can be extracted from agroinfected leaves and used for reverse transcription-polymerase chain reaction (RT-PCR), Northern blot, Western blot, immunoprecipitation, and enzyme assays.
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An Agrobacterium-mediated transient gene expression system for intact leaves
Jyoti Kapila, Riet De Rycke, Marc Van Montagu et al. · Plant Science · 1997 · 727 citations