Difference between revisions of "Part:BBa P0140"
 
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<h2>
 
<h2>
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Tianjin 2021's characterization</h2>
+
Tianjin's 2021 characterization</h2>
 
<b><h3>New information of TetR operon leakage</h3></b>
 
<b><h3>New information of TetR operon leakage</h3></b>
 
<p >Group: Tianjin 2021<br>
 
<p >Group: Tianjin 2021<br>
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Author: Ruiqi Liu<br>
+
Author: Ruiqi Liu, Lingwei Ding<br>
 
Summary: we added information about TetR operon leakage.
 
Summary: we added information about TetR operon leakage.
 
</p>
 
</p>
 
<i><h3>Background</h3></i>
 
<i><h3>Background</h3></i>
  
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[[File:Tianjin-Parts1.png|400px|thumb|right|Figure 1.Design]]
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[[File:Tianjin-Parts1.png|400px|thumb|right|Figure 1.Carotene plasmid with TetR operon]]
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<p>In order to better apply the tetR operon in the project, we characterized the leakage amount of the tetR operon. We constructed a carotene plasmid with tetR operon, and transformed it into yeasts, then cultured the yeasts without Atc inducer. By observing the color of the colony, the leakage of tetR operon can be roughly determined.</p>
+
<p>In our project, we want to use TetR operon to control the expression of mcherry gene. Thus, we want to test whether TetR operon can strictly regulate the expression of gene or not. We constructed a carotene plasmid with TetR operon, and transformed it into Saccharomyces cerevisiae. If the carotene gene controlled by TetR operon leaks, the yeasts will turn orange.</p>
 
<i><h3>Result</h3></i>
 
<i><h3>Result</h3></i>
  
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<p>Our experimental results show that the tetR operon is very easy to leak, and the expression amount of the circuit is different and difficult to test. Therefore, the results clearly indicated that in strict experiments, using the tetR operon alone will affect the experimental results.</p>
+
<p>We cultured the yeasts after transforming the carotene plasmid. We were surprised to find that, before using Atc inducer, a large proportion of yeasts was orange. Our experimental results show that the tetR promoter is very easy to leak, and the expression amount of the circuit is different and difficult to control. Therefore, the other iGEMers should aware that TetR operon is not a good choice if you want to strictly control the expression of structure gene.</p>
  
  
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[[File:Tianjin-Parts2.jpg|300px|thumb|left|Figure 2.Result]]
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[[File:Tianjin-Parts2.jpg|300px|thumb|left|Figure 2.Saccharomyces cerevisiae with carotene plasmids shows that TetR operon is easy to leak ]]
  
  

Latest revision as of 05:24, 21 October 2021


PoPS -> TetR [S0163]


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Tianjin's 2021 characterization

New information of TetR operon leakage

Group: Tianjin 2021
Author: Ruiqi Liu, Lingwei Ding
Summary: we added information about TetR operon leakage.

Background
Figure 1.Carotene plasmid with TetR operon

In our project, we want to use TetR operon to control the expression of mcherry gene. Thus, we want to test whether TetR operon can strictly regulate the expression of gene or not. We constructed a carotene plasmid with TetR operon, and transformed it into Saccharomyces cerevisiae. If the carotene gene controlled by TetR operon leaks, the yeasts will turn orange.

Result

We cultured the yeasts after transforming the carotene plasmid. We were surprised to find that, before using Atc inducer, a large proportion of yeasts was orange. Our experimental results show that the tetR promoter is very easy to leak, and the expression amount of the circuit is different and difficult to control. Therefore, the other iGEMers should aware that TetR operon is not a good choice if you want to strictly control the expression of structure gene.


Figure 2.Saccharomyces cerevisiae with carotene plasmids shows that TetR operon is easy to leak