lec 8 - Activators and repressors of transcription...

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Activators and repressors of transcription Transcriptional control elements like enhancers are binding sites for regulatory proteins. These proteins can be identified by biochemical techniques. DNase I footprinting Electrophoretic mobility shift assay (also called EMSA or „gel shift‟).
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Experimental Methods DNase I footprinting experiments reveal specific binding sites for DNA binding proteins. EMSA or gel shift assays can be used to detect DNA binding proteins during biochemical purification. Co-transfection assays in cultured cells are used to evaluate whether a protein encoded by a known gene is a transcription factor.
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DNase I footprinting: Logic: If a protein is bound to a specific DNA sequence, it will protect that sequence from nuclease digestion. DNA is end-labelled and partially digested with a nuclease that cuts randomly. Is used to map precise binding sites of a protein to DNA.
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