BIMM 100 Lecture Notes - Lecture 11: Deoxyribonuclease, Electrophoretic Mobility Shift Assay, Northern Blot
28 Feb 2018
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Rna polymerase works as helicase to open dsdna and creates initial rna strands: as rna is created, it dissociates from dna to become single stranded. Rna polymerase moves with the transcription: polymerase falls off from dna at stop sight to leave single stranded mrna. How to find all the promoter regions in your genome: extract whole mrna from cells. For a given gene, you know promoter region and coding region: very small sequences in whole promoter region (~10-20bp out of 1kb) are functional elements that are essential for the promoter. 2 types: common: most of genes have these small elements (ex: tata box, gene specific: only exist in small portion of genes. Rna sequence and you can detect using radiolabeled antisense probe. If a smaller region gives the same amount of activity, you know the region that was deleted was not part of the promoter. If on a gel, if only dna present, migrates fast.
