Difference between revisions of "Part:BBa K3221200"
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'''Design: '''
 
'''Design: '''
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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.
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In order to demonstrate the function of GAL1 promoter, we cultured transformed yeast and WT in YPD medium and YPG medium separately. Through detecting green fluorescence both in Fluorescence microscope(Figure.1 (a)) and plate reader(Figure.1 (b)) to verify the function of GAL1 promoter.
 
   
 
   
 
[[File: T--SCU-China--scu-pGALdelay2019.png|450px |]]
 
[[File: T--SCU-China--scu-pGALdelay2019.png|450px |]]
  
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'''Fig.1 Fluorescence observation in yeGFP transformed <i>S.cerevisiae</i> 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. '''  
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'''Figure.1 ''' '''(a) Fluorescence observation in yeGFP transformed <i>S.cerevisiae</i> BY4741. ''' ''' Series 1: BY4741 WT in YPD medium；Series 2: BY4741 WT in YPG medium; Series 3, Series 5: Transformed BY4741 in YPD medium; Series 4, Series 6: Transformed BY4741 in YPG medium. Visible Fluorescence in Series 4 and Series 6. ''' ''' (b) The fluorescence of <i>S.cerevisiae</i>BY4741 transformed pYES2-yeGFP. '''  ''' YPD: YPD medium, YPG: YPG medium, BY4741 WT: <i>S.cerevisiae</i>, BY4741 wildtype, BY4741 transformed: <i>S.cerevisiae</i>, BY4741 transformed plasmid pYES2-yeGFP, excitation light is 488nm and emission light is 525nm. '''
  
 
'''Result & conclusion: '''
 
'''Result & conclusion: '''
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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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Generally, we verified the function of GAL1 promoter by observing green fluorescence in B4(a) and B6(a). In other words, the GAL1 promoter could induce expression of downstream gene (yeGFP in this experiment) in galactose.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 15:48, 20 October 2019


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


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

Design: In order to demonstrate the function of GAL1 promoter, we cultured transformed yeast and WT in YPD medium and YPG medium separately. Through detecting green fluorescence both in Fluorescence microscope(Figure.1 (a)) and plate reader(Figure.1 (b)) to verify the function of GAL1 promoter.

T--SCU-China--scu-pGALdelay2019.png

Figure.1 (a) Fluorescence observation in yeGFP transformed S.cerevisiae BY4741. Series 1: BY4741 WT in YPD medium；Series 2: BY4741 WT in YPG medium; Series 3, Series 5: Transformed BY4741 in YPD medium; Series 4, Series 6: Transformed BY4741 in YPG medium. Visible Fluorescence in Series 4 and Series 6. (b) The fluorescence of S.cerevisiaeBY4741 transformed pYES2-yeGFP. YPD: YPD medium, YPG: YPG medium, BY4741 WT: S.cerevisiae, BY4741 wildtype, BY4741 transformed: S.cerevisiae, BY4741 transformed plasmid pYES2-yeGFP, excitation light is 488nm and emission light is 525nm.

Result & conclusion: Generally, we verified the function of GAL1 promoter by observing green fluorescence in B4(a) and B6(a). In other words, the GAL1 promoter could induce expression of downstream gene (yeGFP in this experiment) in galactose.