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Transcrption factor-promoter (cis-element) interaction

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Transcrption factor-promoter (cis-element) interaction

Postby roniadam » Sun Aug 11, 2013 2:43 am

I want to transfect my cells with a vector for promoter activity analaysis, I need to detect the binding effeciency of some transcription factors to the promoter, what technique should I use? I have read about ChIP but don't know how to design and perform this experiment practically , your help is highly appreciated.
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Postby bravebeaker » Thu Aug 22, 2013 3:31 am

are you testing in plant or animal cells.
I have been working with plant cells and the most common and easy way I guess is the reporter gene assay.
the promoter in question is designed in a vector carrying the GUS reporter gene then you can measure GUS activity for instance...
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Postby JackBean » Thu Aug 22, 2013 11:43 am

I'd say roniadam wants to measure exactly the binding.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Postby bravebeaker » Mon Aug 26, 2013 5:45 am

How about EMSA then? electromobility shift assay also applicable for the same purpose more or less...
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