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Transient Expression Assay by Agroinfiltration of Leaves

  • Min Woo Lee
  • Yinong Yang
Part of the Methods in Molecular Biology™ book series (MIMB, volume 323)

Abstract

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.

Key Words

Transient gene expression Agrobacterium tumefaciens agroinfiltration GUS staining dexamethasone-inducible expression 

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Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Min Woo Lee
    • 1
  • Yinong Yang
    • 2
  1. 1.Department of Molecular Genetics and Cell BiologyUniversity of ChicagoChicago
  2. 2.Department of Plant PathologyUniversity of ArkansasFayetteville

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