Difference between revisions of "Part:BBa K2368006"

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<partinfo>BBa_K2368006 short</partinfo>
 
<partinfo>BBa_K2368006 short</partinfo>
  
<h4>General</h4>
+
<h3>General</h3>
  
<p>This composite part is based on the Saccharomyces cerevisiae pheromone signaling pathway. It contains a yeast induced promoter Pfus, a RFP reporter, and a yeast terminator. Pfus is activated by &#945; pheromone, thereby initiating the expression of the reporter gene.</p>
+
<p>To measure the sweetness of sweeteners, we designed the detection line. This composite part is based on the Saccharomyces cerevisiae pheromone signaling pathway. It contains a yeast induced promoter Pfus, a mRFP reporter, and a yeast terminator. In CEN.PK2-1C(a mating type yeast) Pfus is activated by α pheromone, thereby initiating the expression of the reporter gene.</p>
  
 
<p>there will be a picture here.</p>
 
<p>there will be a picture here.</p>
  
<h4>Our experiment</h4>
+
<h3>Our experiment</h3>
  
 
<p>We constructed Pfus-RFP-CYC1t-PRS42K recombinant plasmid, and transformed it into CENPK2-1C. Cells transformed with the signal reporter device were cultured to the exponential phase and treated with &#945; pheromone.</p>
 
<p>We constructed Pfus-RFP-CYC1t-PRS42K recombinant plasmid, and transformed it into CENPK2-1C. Cells transformed with the signal reporter device were cultured to the exponential phase and treated with &#945; pheromone.</p>

Revision as of 02:18, 14 October 2017


Introduction

Signal reporter device (Pfus-mRFP-CYC1t)

General

To measure the sweetness of sweeteners, we designed the detection line. This composite part is based on the Saccharomyces cerevisiae pheromone signaling pathway. It contains a yeast induced promoter Pfus, a mRFP reporter, and a yeast terminator. In CEN.PK2-1C(a mating type yeast) Pfus is activated by α pheromone, thereby initiating the expression of the reporter gene.

there will be a picture here.

Our experiment

We constructed Pfus-RFP-CYC1t-PRS42K recombinant plasmid, and transformed it into CENPK2-1C. Cells transformed with the signal reporter device were cultured to the exponential phase and treated with α pheromone.

We added 10umol/L α pheromone into the transformed yeast. And after 4 hours, it was fluorescent.

there will be a picture here.

In order to characterize the function of signal reporter device, we measured absorbance and fluorescence of the cells in the plate reader after treating them with varying concentrations of α pheromone.

there will be a picture here.

As shown in this figure, adding higher concentration of α pheromone, the fluorescence intensity of the yeast increases. After 20 hours, α pheromone gradually failed, and the fluorescence intensity also decreased.

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 993
    Illegal AgeI site found at 1105
  • 1000
    COMPATIBLE WITH RFC[1000]