Difference between revisions of "Part:BBa P0140"

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

@@ Line 13: / Line 13: @@
-<partinfo>TetR BBa_P0140</partinfo>
+<h2>
-<b><h2>Tianjin 2021's characterization</h2></b>
+Tianjin's 2021 characterization</h2>
-<b><h3>New information of TetR promoter leakage</h3></b>
+<b><h3>New information of TetR operon leakage</h3></b>
 <p >Group: Tianjin 2021<br>
-Author: Ruiqi Liu<br>
+Author: Ruiqi Liu, Lingwei Ding<br>
-Summary: we added information about TetR promoter leakage.
+Summary: we added information about TetR operon leakage.
 </p>
 <i><h3>Background</h3></i>
-[[File:Tianjin-Parts1.png|400px|thumb|right|Figure 1.Design]]
+[[File:Tianjin-Parts1.png|400px|thumb|right|Figure 1.Carotene plasmid with TetR operon]]
-<p>In order to better apply the TetR promoter in the project, we characterized the leakage amount of the TetR promoter. We constructed a carotene plasmid with TetR promoter, and transformed it into yeasts, then cultured the yeasts without AHT inducer. By observing the color of the colony, the leakage of TetR promoter 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>
-<p>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 test. Therefore, the results clearly indicated that in strict experiments, using the TetR promoter 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>
-<h3><i>Design</i></h3>
+[[File:Tianjin-Parts2.jpg|300px|thumb|left|Figure 2.Saccharomyces cerevisiae with carotene plasmids shows that TetR operon is easy to leak ]]
-[[File:Tianjin-Parts2.jpg|300px|thumb|left|Figure 2.Result]]