BIOL 200 Lecture Notes - Lecture 9: Dna-Binding Domain, Elution, Sv40

• School: McGill University
• Department: Biology
• Course: BIOL 200
• Professor: Kenneth Hastings

Document Summary

All genes use gtfs but some genes use transcription factors. Emsa or gel shift assay can be used to detect dna binding proteins during biochemical puri cations. Dnase i foot printing experiments: reveals speci c binding sites and can be used as an assay. Segment of dna bound to protein will migrate slower in gel than dna that is alone. Better than footprint for quantitive analysis (for mutant gal4 bind to uas) Find out which proteins bind to speci c enhancer regions. To separate proteins > liquid chromatography: separation can be based on mass, charge, or af nity to antibodies. If protein is bound to a speci c sequence, it will protect that sequence from nuclease digestion. Partially digested with low conc. of nuclease that cuts randomly. Used to map praise site of a protein to dna. Tbp binds to -40 to -15 regions. Map the binding site by footprint experiments. Synthesize dna sequence contain multiple copies of binding site.