MCELLBI C148 Lecture Notes - Lecture 13: Regulon, Rna-Seq, Repressor Lexa
3 May 2020
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Review: regulation of transcription initiation: most genes are regulated at the level of transcription initiation. In general, a higher rate of transcription initiation (= more mrna molecules) leads to more protein. Review: how are dna repair genes regulated: make a reporter fusion, transposon mutagenesis, screen for higher uninduced expression and/or lower induced expression, map transposon insertions, genetic analysis to identify regulators. The dna damage response (sos response) is an example of a regulon: regulon = group of genes controlled by the same regulatory pathway. Differences in different number of reads are differentially expressed! Mmc the gene is not expressed but in the mmc it is expressed, so difference in blue color. Strategy 2: genetic screen: mutagenize with a transposon that can make transcriptional fusions to lacz, replica plate onto plates with and without fungal extract, screen for and isolate strains with differences in blue color, map transposon insertions. Validation for both: construct/re-test reporter fusions or qpcr.
