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Lecture 13

BIO372 - Lecture 13.pdf

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Gordon Anderson

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Lecture 13 March-04-14 10:14 AM Transcriptionoverview 1. Search DNA for promoters,polymerasebinds to promoters.If successful, DNA opens up (acts like helicase) 2. These are called board of transcripts, they end up in short fragments and get ejected 3. Only few are successful to go into elongation 4. Termination process occurs DNA is the template for transcription Bubble is moving in the opposite direction, nucleotide is added in the 3' end Nontemplate strand is the coding strand RNA chain growth is 5’ to 3’ Prokaryotic promoters Pribnow box at -10 box Notice the way letters are written (some are written in small/ capital letters) they represent the frequency - commonin DNA motifs Any given promoterdiffers in one or morebases. If the promotermatches, it is a very strong promoter How are promotersdiscovered? - We can look at areas protected by RNA polymerase - We can look for regions of commonality - this can gives us candidates to be looked over in other experiments - We can look at transcriptional outputs (change nucleotide one at a time and comparethe activity) There is an optimum distance between two boxes Prokaryotic promoters Holoenzymeand core enzyme has catalytic activity Sigma factor allows specific binding and finding of promoters,this is important,only certain strands can bind We label DNA and flow mixture into nitrocellulose, if DNA is not bound, its gonna flow through The core has low affinity to DNA template If there is sigma factor, complex is much morestable because there is moreenzymes To look at stability, look at half life of binding of DNA and protein The carboxy terminal domain (CTD) of a binds to the UP element of a promoter. Carboxy terminus binds to DNA (UP site), N-terminus binds to enzyme Overview of transcription initiation Initiation seem to be wasteful Included: E.coli RNA pol; a strong promoter; and heparin (a
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