Promoter, Chip, transcription factors etc.
Posted 21 July 2009 - 11:44 AM
Posted 21 July 2009 - 02:00 PM
If you know the upregulated protein, do its promoter analysis through bioinformatics. (TF binding sites, evolutionary conservation thereof etc).
Hi everybody! I have a mouse strain lacking a gene. In tissues lysate derived from this mouse, there is a protein with increased expression compared to the wild type. When I add a chemical that compensate for the lack of the gene, the expression of the protein is downregulated. I am eager to know which transcription factor regulates an expression of the protein, but don't know how to do it. If I would wanted to check whether a given transcription factor binds to some promoter(s), I'd do Chip employing antibody against transription factor followed by PCR, but what to do in such sort of reversed situation? Many-many thanks for ideas! (I'll mention your help if get published in Nature )
Then, make an intelligent and informed choice of transcription factors that you think may really be involved in the pathway, and do ChIP, reporter analysis and EMSAs for it.