Identify txn factors?? - (Apr/30/2010 )
Hi Guys! So I'm wondering if anyone knows if there's a 'final say' assay on what is or is not a transcription factor?
Basically, I'm trying to design a project and have a protein I THINK is a transcriptional regulator, because it's homolog is a txn factor, and some of the proteins it binds to are also txn regulators. I'm already planning to do a microarray to look at gene expression, followed by EMSA to see if it binds DNA in promoter regions of any genes that I get from the microarray.
If I have candidate genes, is the luciferase reporter assay the commonly used experiment to definitely say 'this is a txn factor'?
I'm not familiar with the luciferase assay. I gather that I put the promoter region of a candidate gene in front of the luciferase gene. When I transfect it in, with a plasmid for my protein, I look to see if transcript levels change? What if my protein requires co-activators?
Any input would be awesome. Thanks a bunch!!
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