Greetings to all,
i want to study the role of a transcriptional co-activator in the regulation of a certain gene expression, through binding to specific transcription factors. In other words, i would like to evaluate if the coactivator of interest can bind to a transcription factor that in turn binds to specific DNA regions and regulates a particular gene of interest. Since i am completely inexperienced in this field can anyone suggest techniques to do so or any research papers with the same logic? Thank you in advance.
Hello, Theodoros.
The followings would guide you. References and cited papers therein.
doi: 10.1101/cshperspect.a000034
DOI 10.1016/j.cmet.2005.05.004
doi: 10.1016/j.tcb.2012.04.002
Regards.
Dear Theodoros,
Greeting
For studying the protein interactions, the yeast two-hybrid approach is powerful tool. you can define you desired protein as bait, and then checking their pairwise interaction to other proteins as preys.
Please have look the below papers:
10.1101/pdb.prot5431.
10.3390/ijms10062763
I would suggest you to perform Chromatine immunoprecipitation (ChIP). Using this technique you can purify DNA (chromatine) and immunoprecipitate your transcription factor using antibody then immunoblot for your protein of interest (which you believe is binding to the transcription factor).
You can actually play around with this to address this question using multiple strategies. There many reports using ChIP please read about them
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