Composite

Part:BBa_K3221200

Designed by: Yitao Ke   Group: iGEM19_SCU-China   (2019-10-04)
Revision as of 14:27, 15 October 2019 by Koursera (Talk | contribs)


Galactose-mediated induction system, 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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1199


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.

T--SCU-China--scu-pGAldelay2019.png

Fig.1 Fluorescence observation A1 and B1: WT in YPG;A2 and B2 : Transformed in YPD;A3, A4, B3 and B4: Transformed in YPG. Visible Fluorescence in B3 and B4.

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.


[edit]
Categories
Parameters
None