CMMB 461 Lecture Notes - Lecture 24: Power Law, Lac Operon, Histidine

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In chip-chip, we"re tagging transcription factors (protein centered) Promoter in this assay is part of the gene. Query is not the transcription factor, you"re not looking for the transcription factor targets. In this, you"re looking for transcription factors that bind to the promoter, not promoters that bind to the transcription factor. Procedure: make a library (clone in front of strong activation domain), from there, take entire library (transcription factor library, or orfeome) and transform into budding yeast cell. Yeast cell has promoter of interest (studying) hooked to the reporter. This organism is normally his minus (his-), meaning that it cannot grow in the absence of histidine (prototroph). If transcription factor binds to promoter and that fires up the reporter, expect yeast cells to be blue in colour and able to grow in the absence of histidine. If transcription factor binds to promoter, reporter gene is activated.

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