Difference between revisions of "Part:BBa K3221200"
Line 5: Line 5:
 
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.  
 
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.  
  
−
Then, we designed to culture transformed yeast and WT in YPD medium and YPG medium separately. Through detecting green fluorescence to verify the function of GAL1 promoter.
 
−
 
−
[[File: T--SCU-China--scu-pGAldelay2019.png|450px|]]
 
−
 
−
'''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.
 
−
 
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
Line 20: Line 12:
 
<partinfo>BBa_K3221999 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3221999 SequenceAndFeatures</partinfo>
  
 +
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.
 +
 +
[[File: T--SCU-China--scu-pGAldelay2019.png|450px|]]
 +
 +
'''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.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 14:27, 15 October 2019


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.