Difference between revisions of "Team:NEU China B/Improve"

 
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<H2>
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<h2>BBa_k2824006:T7-lldPRD operon promoter-GFP</h2>
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BBa_k2824008:Lldr-T7-lldPRD operon promoter-GFP
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<div class="p">
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</H2>
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LLDPRD promoter can sense the presence of lactic acid, in the absence of lactic acid, only a lower background level
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<div class="p">
+
of expression, but in the presence of lactic acid can open gene expression. We integrated the GFP reporter gene
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<br>The role of this part is to try to control the concentration of lactic acid on the LLDPRD promoter under the
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into downstream this promoter to respond to the presence of lactic acid. In order to improve the GFP expression, we
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control of quorum sensingg. In the case of LLdr gene expression, the subsequent gene expression was initiated.
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integrated a strong promoter, T7, into the upstream of lldPRD region.
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<br>This part also is constructed based on the part lldRO1-plldR-lldRO2 (BBa_k82000) . lldRO1-plldR-lldRO2 part is a
+
</div>
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lactic acid regulated promoter that is open to gene transcription in the presence of lactic acid. Comparing with the
+
<div class="p">
−
T7-lldPRD operon promoter-GFP part, the improvement of this part is that the LLDR is added to part and placed under
+
This part is constructed based on the part lldRO1-plldR-lldRO2 (BBa_k82000) . lldRO1-plldR-lldRO2 part is a lactic
−
the control of two promoters respectively to form a double expression regulatory system. This comparison with the two
+
acid regulated promoter that is open to gene transcription in the presence of lactic acid. It can effectively
−
plasmids working together is more convenient for testing the success of the system.
+
improve the expression of the target gene and detect whether the target gene is expressed or not. At the same time,
−
<br>We can easily find that the Lldr-T7-lldPRD operon promoter-GFP part are in a higher level of expression comparing
+
it can accurately determine the amount of the target gene. This allows the overall work of our system to be
−
to the lldPRD operon promoter-GFP when the lactate concentration is lower than nearly 1.5m mol/L. Considering that
+
reflected in digital form.
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the lactate concentration in yogurt is generally no more than 1m mol/L, the conclusion can given that the T7 promoter
+
</div>
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can enhance the signal intensity for our experiments.
+
<h2>
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<div>
+
Improve the Characterization of BBa_k82000
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+
</h2>
−
<h2>
+
<div class="p">
−
BBa_k2824006:T7-lldPRD operon promoter-GFP
+
In the following figure, we compared the expression of lldPRD operon promoter-GFP and T7-lldPRD operon
−
</h2>
+
promoter-GFP. The T7 promoter could enhance gene expression when the concentration of lactic acid was less than 1 m
−
<div class="p">
+
mol/L. Considering that the lactate concentration in yogurt is generally no more than 1m mol/L, the conclusion can
−
<br>LLDPRD can sense the presence of lactic acid, in the absence of lactic acid, only a lower background level of
+
given that the T7 promoter can enhance the signal intensity for our experiments (Figure 1).
−
expression, but in the presence of lactic acid can open expression, gene transcription. We added the GFP gene into
+
</div>
−
the gene and used green fluorescence as a marker to recognize the presence of lactic acid. In addition, as a strong
+
 
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promoter, T7 can improve the expression level of the target gene.
+
Figure 1:lldPRD operon promoter-GFP vs T7-lldPRD operon promoter-GFP
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<img src="https://static.igem.org/mediawiki/2018/e/ea/T--NEU_China_B--ct8.png">
+
 
−
<img src="https://static.igem.org/mediawiki/2018/3/34/T--NEU_China_B--improve2.png">
+
<h2>
−
<br>This part is constructed based on the part lldRO1-plldR-lldRO2 (BBa_k82000) . lldRO1-plldR-lldRO2 part is a
+
BBa_k2824008:Lldr-T7-lldPRD operon promoter-GFP
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lactic acid regulated promoter that is open to gene transcription in the presence of lactic acid. We added a sequence
+
</h2>
−
of T 7 promoter and GFP fluorescent protein gene before and after this part Separately. It can effectively improve
+
<div class="p">
−
the expression of the target gene and detect whether the target gene is expressed or not. At the same time, it can
+
The aim of this part is using llDPRD promoter to activate downstream GFP reporter gene expression under different
−
accurately determine the amount of the target gene. This allows the overall work of our system to be reflected in
+
lactic acid concentration inductions and this operon is repressed without lactic acid induction. Briefly, LLdR
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digital form.
+
repression protein specifically binds to llDPRD promoter and impedes downstream gene expression, while lactic acid
−
<br>In the following figure, we compared the expression of lldPRD operon promoter-GFP and T7-lldPRD operon
+
enables to antagonize and replace LLdR protein to activate llDPRD promoter (Figure 1). In order to improve the GFP
−
promoter-GFP. The T7 promoter could enhance gene expression when the concentration of lactic acid was less than 1 m
+
expression, we integrated a strong promoter, T7, into the upstream of lldPRD region.
−
mol/L. Considering that the lactate concentration in yogurt is generally no more than 1m mol/L, the conclusion can
+
</div>
−
given that the T7 promoter can enhance the signal intensity for our experiments
+
<div class="p">
−
<img src="https://static.igem.org/mediawiki/2018/3/33/T--NEU_China_B--improve3.png">
+
This part is constructed based on the part lldRO1-plldR-lldRO2 (BBa_k82000). lldRO1-plldR-lldRO2 part is a lactic
−
<img src="https://static.igem.org/mediawiki/2018/c/c6/T--NEU_China_B--improve4.png">
+
acid regulated promoter that is open to gene transcription in the presence of lactic acid. Comparing with the
−
<img src="https://static.igem.org/mediawiki/2018/8/8d/T--NEU_China_B--improve5.png">
+
T7-lldPRD operon promoter-GFP part, the improvement of this part is that the LLDR is added to part and placed under
−
<div>
+
the control of two promoters respectively to form a double expression regulatory system. This comparison with the
−
</html>
+
two plasmids working together is more convenient for testing the success of the system.
 +
</div>
 +
<h2>
 +
Improve the Characterization of BBa_k82000
 +
</h2>
 +
<div class="p">
 +
We can easily find that the Lldr-T7-lldPRD operon promoter-GFP part are in a higher level of expression comparing
 +
to the lldPRD operon promoter-GFP when the lactate concentration is lower than nearly 1.5 mM. Considering that the
 +
lactate concentration in yogurt is generally no more than 1 mM, the conclusion can give that the T7 promoter can
 +
enhance the signal intensity for our experiments (Figure 2).
 +
</div>
 +
<div class="text-center">
 +
<div>
 +
<img src="https://static.igem.org/mediawiki/2018/8/8d/T--NEU_China_B--8end0.png"class="center-block" alt="">
 +
</div>
 +
Figure 1:Lldr-T7-lldPRD operon promoter-GFP
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</div>
 +
<div class="text-center">
 +
<div>
 +
<img src="https://static.igem.org/mediawiki/2018/1/17/T--NEU_China_B--8end1.png" alt="" class="center-block">
 +
</div>
 +
Figure 2:lldPRD operon promoter-GFP vs Lldr-T7-lldPRD operon promoter-GFP
 +
</div>
 +
<div class="p">
 +
To see more details about the construction and result, click the hyperlink below:
 +
<br>
 +
<a href="http://parts.igem.org/Part:BBa_K822000">lldPRD operon promoter + RBS from E. coli ( lldRO1-plldR-lldRO2
 +
)：BBa_k82000</a>
 +
</div>

Latest revision as of 20:55, 17 October 2018

Ruby - Responsive Corporate Tempalte

BBa_k2824006:T7-lldPRD operon promoter-GFP

LLDPRD promoter can sense the presence of lactic acid, in the absence of lactic acid, only a lower background level of expression, but in the presence of lactic acid can open gene expression. We integrated the GFP reporter gene into downstream this promoter to respond to the presence of lactic acid. In order to improve the GFP expression, we integrated a strong promoter, T7, into the upstream of lldPRD region.
This part is constructed based on the part lldRO1-plldR-lldRO2 (BBa_k82000) . lldRO1-plldR-lldRO2 part is a lactic acid regulated promoter that is open to gene transcription in the presence of lactic acid. It can effectively improve the expression of the target gene and detect whether the target gene is expressed or not. At the same time, it can accurately determine the amount of the target gene. This allows the overall work of our system to be reflected in digital form.

Improve the Characterization of BBa_k82000

In the following figure, we compared the expression of lldPRD operon promoter-GFP and T7-lldPRD operon promoter-GFP. The T7 promoter could enhance gene expression when the concentration of lactic acid was less than 1 m mol/L. Considering that the lactate concentration in yogurt is generally no more than 1m mol/L, the conclusion can given that the T7 promoter can enhance the signal intensity for our experiments (Figure 1).
Figure 1:lldPRD operon promoter-GFP vs T7-lldPRD operon promoter-GFP

BBa_k2824008:Lldr-T7-lldPRD operon promoter-GFP

The aim of this part is using llDPRD promoter to activate downstream GFP reporter gene expression under different lactic acid concentration inductions and this operon is repressed without lactic acid induction. Briefly, LLdR repression protein specifically binds to llDPRD promoter and impedes downstream gene expression, while lactic acid enables to antagonize and replace LLdR protein to activate llDPRD promoter (Figure 1). In order to improve the GFP expression, we integrated a strong promoter, T7, into the upstream of lldPRD region.
This part is constructed based on the part lldRO1-plldR-lldRO2 (BBa_k82000). lldRO1-plldR-lldRO2 part is a lactic acid regulated promoter that is open to gene transcription in the presence of lactic acid. Comparing with the T7-lldPRD operon promoter-GFP part, the improvement of this part is that the LLDR is added to part and placed under the control of two promoters respectively to form a double expression regulatory system. This comparison with the two plasmids working together is more convenient for testing the success of the system.

Improve the Characterization of BBa_k82000

We can easily find that the Lldr-T7-lldPRD operon promoter-GFP part are in a higher level of expression comparing to the lldPRD operon promoter-GFP when the lactate concentration is lower than nearly 1.5 mM. Considering that the lactate concentration in yogurt is generally no more than 1 mM, the conclusion can give that the T7 promoter can enhance the signal intensity for our experiments (Figure 2).
Figure 1:Lldr-T7-lldPRD operon promoter-GFP
Figure 2:lldPRD operon promoter-GFP vs Lldr-T7-lldPRD operon promoter-GFP
To see more details about the construction and result, click the hyperlink below:
lldPRD operon promoter + RBS from E. coli ( lldRO1-plldR-lldRO2 )：BBa_k82000