Difference between revisions of "Part:BBa K2368006"
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<p>We added 5umol/L α pheromone into the transformed yeast. And after 4 hours, it was fluorescent.</p> | <p>We added 5umol/L α pheromone into the transformed yeast. And after 4 hours, it was fluorescent.</p> | ||
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<li style="display: inline-block;"> [[File:T_BIT-China_2017part_10.png|center|center|220px|a]] </li> | <li style="display: inline-block;"> [[File:T_BIT-China_2017part_10.png|center|center|220px|a]] </li> | ||
<li style="display: inline-block;"> [[File:T_BIT-China_2017part_11.png|center|center|220px|b]] </li> | <li style="display: inline-block;"> [[File:T_BIT-China_2017part_11.png|center|center|220px|b]] </li> | ||
</ul></div> | </ul></div> | ||
| − | <p>Fig.3 Images of yeast fluorescence. a: Initial fluorescence image; b: Fluorescence image after 4 hours. </p> | + | <p style="text-align: center">Fig.3 Images of yeast fluorescence. a: Initial fluorescence image; b: Fluorescence image after 4 hours.</p> |
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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> | <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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<h2>Sequence and Features</h2> | <h2>Sequence and Features</h2> | ||
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Latest revision as of 07:21, 24 October 2017
Introduction
Signal reporter device (Pfus-mRFP-CYC1t)
General
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.
Fig.1 The detection circuit
We constructed Pfus-mRFP-cyc1t-PRS42K recombinant plasmid, and transformed it into CEN.PK2-1C (a type).
Fig.2 The result of colony PCR
Cells transformed with the signal reporter device were cultured to the exponential phase and treated with α pheromone.
We added 5umol/L α pheromone into the transformed yeast. And after 4 hours, it was fluorescent.
Fig.3 Images of yeast fluorescence. a: Initial fluorescence image; b: Fluorescence image after 4 hours.
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.
Fig.4 mRFP intensity of CEN.PK2-1C induced by α factor
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 993
Illegal AgeI site found at 1105 - 1000COMPATIBLE WITH RFC[1000]