Part:BBa_K3221200
Galactose-mediated induction expression system for S.cerevisiae, yeGFP as a reporter
GAL system is a galactose-mediated induction system in yeast. To be specific, GAL1 promoter can induce downstream gene expression under the control of GAL4 protein. What’s more, in order to simplify the verification of GAL1 promoter function, we chose the yeGFP as a reporter instead of Cns1 and Cns2 in our project.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 150
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1199
Design: In order to demonstrate our design, we cultured transformed yeast and WT in YPD medium and YPG medium separately. Through detecting green fluorescence to verify the function of GAL1 promoter.
Fig.1 Fluorescence observation in yeGFP transformed S.cerevisiae BY4741. Series 1: BY4741 WT in YPG medium；Series 2: Transformed BY4741 in YPD medium；Series 3 and Series 4: Transformed BY4741 in YPG medium. Visible Fluorescence in Series3 and Series 4.
Result & conclusion: Generally, we verified the function of GAL1 promoter by observing green fluorescence in B3(Fig.1) and B4(Fig.1). In other words, the GAL1 promoter could induce expression of downstream gene (yeGFP in this experiment) in galactose.
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