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Transient Expression Assay by Agroinfiltration of Leaves

 Last Update: 2020-11-20
 Source: Internet
 Author: User

Tags

reverse transcriptase polymerase chain reaction

gene

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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 β-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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