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

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

Revision as of 20:55, 17 October 2018

Ruby - Responsive Corporate Tempalte

Figure 1:Lldr-T7-lldPRD operon promoter-GFP
Figure 2:lldPRD operon promoter-GFP vs Lldr-T7-lldPRD operon promoter-GFP