CSB349H1 Lecture Notes - Lecture 4: Sea Urchin, In Situ Hybridization, Chromosome
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Promoters: in eukaryotic promoters, there"s two major parts. Not as close as the core to the transcription start site: experiment. Depending on which plasmid that drives which reporter gene expression, they could find the key elements in non-coding dna. You have a piece of dna that you don"t know what it does. You start deleting pieces of it and see if you can still see reporter gene expression. When you can"t see the expression, it means that the part of dna is important. You have to test the pieces one by one. You take copies of your promoter and you label one end of them typically with a radioactive label. Then you do a digestion with dnase1 (will cut the dna randomly) You get fragments with the label of all sizes. You run the fragments on the gel and you see many different sizes. If you add proteins before doing the digestion, it will protect the.