What is transactivation assay?

The principle of in vitro androgen transactivation assay, based on stable transfection of a cell line with two plasmids; one encoding the androgen receptor and the other, the androgen response element (ARE) upstream of a reporter (REP) gene such as luciferase.

What is transactivation activity?

From Wikipedia, the free encyclopedia. In the context of gene regulation: transactivation is the increased rate of gene expression triggered either by biological processes or by artificial means, through the expression of an intermediate transactivator protein.

What is a specific Transactivator?

Transactivators bind specific DNA sequences at open enhancers or promoters and recruit co-activators that contact the basal transcription machinery to effect transcription.

Can a transactivation assay be used in plants?

Although a similar system in a yeast system can also be used, it poses a problem in that the TF that activates in yeast does not always activate in plants. To detect the possible interaction of TFs, in vivo yeast two‐hybrid and in vitro pull‐down assays can be used.

How is Y1H used in the yeast assay?

Y1H relies on the general principles of the yeast two-hybrid assay. In short, mammalian proteins are exogenously expressed in yeast and their interactions in vivo are measured by the downstream activation of reporter gene constructs.

How is transactivation assay used to screen for TF?

A series of fusions of the GAL4 activation domain and different portions of TF were designed and assayed for their ability to bind to the cis element and activate the HIS3 and lacZ reporter genes in the yeast reporter strain used for screening of the TF.

What are the bait sequences in yeast assay?

1) A DNA bait sequence. This can be a defined promoter sequence, another defined cis-acting sequence, regulatory sequences from exons, introns, 5’ or 3’-noncoding regions, or even centromere or telomere sequences. 2) Bait plasmid.