Pulsed Induction of Circadian Clock Genes in Arabidopsis Seedlings
pmid: 24792053
Pulsed Induction of Circadian Clock Genes in Arabidopsis Seedlings
The Alc-inducible system is a simple, yet effective, "gene switch" that can be used to transiently induce gene expression in Arabidopsis. Here we provide a protocol for using the Alc-inducible system to give a pulse in expression of a circadian clock gene in transgenic seedlings. The line we use as an example harbors an Alc-inducible copy of the CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) gene (Alc∷CCA1). Alc∷CCA1 seedlings are grown on solid MS medium and subsequently treated with ethanol vapor. Because the ethanol is quickly absorbed into the medium upon exposure, the seedlings are moved to fresh plates following treatment to avoid continuous induction. After the induction, the seedlings are harvested over a time-course for future total RNA and/or protein extraction that can be used for subsequent gene expression analyses.
- University of California, San Francisco United States
- UCLA Clinical and Translational Science Institute United States
Arabidopsis Proteins, Gene Expression Regulation, Plant, Seedlings, Alcohols, Circadian Clocks, Arabidopsis, Promoter Regions, Genetic
Arabidopsis Proteins, Gene Expression Regulation, Plant, Seedlings, Alcohols, Circadian Clocks, Arabidopsis, Promoter Regions, Genetic
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