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| − | __NOTOC__
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| − | <h1>Introduction</h1>
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| − | <partinfo>BBa_K2368006 short</partinfo>
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| − | <h3>General</h3>
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| − | <p>To measure the sweetness of sweeteners, we designed the detection circuit. 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>
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| − | [[File:T_BIT-China_2017part_8.png|center|500px|默认文字]]
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| − | <p style="text-align: center">fig.1 The detection circuit</p>
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| − | <p>We constructed Pfus-mRFP-cyc1t-PRS42K recombinant plasmid, and transformed it into CEN.PK2-1C (a type). </p>
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| − | [[File:T_BIT-China_2017part_9.png|center|300px|默认文字]]
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| − | <p style="text-align: center">fig.2 The result of cPCR</p>
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| − | <p>Cells transformed with the signal reporter device were cultured to the exponential phase and treated with α pheromone.</p>
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| − | <p>We added 5umol/L α pheromone into the transformed yeast. And after 4 hours, it was fluorescent.</p>
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| − | <div><ul>
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| − | <li style="display: inline-block;"> [[File:T_BIT-China_2017part_10.png|thumb|none|220px|a]] </li>
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| − | <li style="display: inline-block;"> [[File:T_BIT-China_2017part_11.png|thumb|none|220px|b]] </li>
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| − | </ul></div>
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| − | <p>In order to characterize the function of detection circuit, we measured absorbance and fluorescence of the cells in the plate reader after treating them with varying concentrations of α pheromone. </p>
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| − | [[File:T_BIT-China_2017part_12.png|center|500px|默认文字]]
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| − | <p style="text-align: center">fig.4 RFP intensity of CEN.PK2-1C induced by α factor</p>
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| − | <p>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.</p>
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| − | <!-- Add more about the biology of this part here
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| − | ===Usage and Biology===
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| − | <h2>Sequence and Features</h2>
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| − | <partinfo>BBa_K2368006 SequenceAndFeatures</partinfo>
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| − | <!-- Uncomment this to enable Functional Parameter display
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| − | ===Functional Parameters===
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| − | <partinfo>BBa_K2368006 parameters</partinfo>
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