Difference between revisions of "Part:BBa K4307021"

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!--PnisRRH07 --
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<partinfo>BBa_K4307021 short</partinfo>
  
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This part is derived from PnisA, which is originated from <i>Lactococcus lactis subsp. Lactis</i>, by the improvement done by our own software tool to enhance the promoter.
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partinfoBBa_K4307021 shortpartinfo
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This part is derived from PnisA, which is originated from iLactococcus lactis subsp. Lactisi, by the improvement done by our own software tool to enhance the promoter.
 
  
  
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<!-- Add more about the biology of this part here
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!-- Add more about the biology of this part here
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===Usage and Biology===
 
===Usage and Biology===
  
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span class='h3bb'Sequence and Featuresspan
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<h2>Sequence and Features</h2>
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partinfoBBa_K4307021 SequenceAndFeaturespartinfo
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<partinfo>BBa_K4307021 SequenceAndFeatures</partinfo>
  
  
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!-- Uncomment this to enable Functional Parameter display  
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<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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partinfoBBa_K4307000 parameterspartinfo
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<partinfo>BBa_K4307000 parameters</partinfo>
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<h2>Characterization </h2>
  
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h2Characterization h2
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<p>The following figure demonstrates our successful construction.</p>
  
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pThe following figure demonstrates our successful construction.p
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<br>
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<div class="center">
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    <div class="thumb tnone">
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        <div class="thumbinner" style="width:50%;">
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            <a href="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-gel.png" class="image">
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                <img alt="" src="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-gel.png" width="100%" height=auto class="thumbimage" /></a>                  <div class="thumbcaption">
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                <div class="magnify">
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                    <a href="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-gel.png" class="internal" title="Enlarge"></a>
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                </div>
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                <b>Figure 1: </b> <b>The construction results of PnisRRH07.</b>
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            </div>
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        </div>
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    </div>
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</div>
  
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<h3>Fluorescence assay was done to characterize the bio brick. </h3>
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httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307021-gel.pngbr
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<p>We introduced these 10 promoters into our system into MACH1-T1 strain to detect EGFP fluorescence signal with and without nisin induction by microplate reader. Results demonstrated that fluorescence signals were enhanced both with and without nisin induction except for PnisRRH07 and PnisRRH10(Figure 2).  </p>
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                 bFigure 1 b bThe construction results of PnisRRH07.b
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<p>
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  <br>
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<div class="center">
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    <div class="thumb tnone">
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        <div class="thumbinner" style="width:50%;">
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            <a href="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-1.png" class="image">
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                 <img alt="" src="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-1.png" width="100%" height=auto class="thumbimage" /></a>                  <div class="thumbcaption">
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                <div class="magnify">
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                    <a href="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-1.png" class="internal" title="Enlarge"></a>
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                </div>
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                <b>Figure 2: </b> <b> Fluorescence spectrophotometry results of the 10 mutant promoters with the highest predictive strength.</b>
 
                  
 
                  
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h3Fluorescence assay was done to characterize the bio brick. h3
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pWe introduced these 10 promoters into our system into MACH1-T1 strain to detect EGFP fluorescence signal with and without nisin induction by microplate reader. Results demonstrated that fluorescence signals were enhanced both with and without nisin induction except for PnisRRH07 and PnisRRH10(Figure 2).  p
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</div>
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  br
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httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307021-1.pngbr
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                bFigure 2 b b Fluorescence spectrophotometry results of the 10 mutant promoters with the highest predictive strength.b
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Furthermore, flow cytometry results confirmed that percentages of positive fluorescence signal bacteria also increased both with and without nisin induction(Figure 3), indicating that the improvement of promoter PnisA by our software tool was practical.  
 
Furthermore, flow cytometry results confirmed that percentages of positive fluorescence signal bacteria also increased both with and without nisin induction(Figure 3), indicating that the improvement of promoter PnisA by our software tool was practical.  
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br
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<br>
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httpsstatic.igem.wikiteams4307wikiwiki-parts-filesmainpagebba-k4307015-2.pngbr
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<div class="center">
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                 bFigure 3 b b Flow cytometry results of the 10 mutant promoters with the highest predictive strength. b
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    <div class="thumb tnone">
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        <div class="thumbinner" style="width:50%;">
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            <a href="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307015-2.png" class="image">
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                 <img alt="" src="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-2.png" width="100%" height=auto class="thumbimage" /></a>                  <div class="thumbcaption">
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p
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                <div class="magnify">
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                    <a href="https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307021-2.png" class="internal" title="Enlarge"></a>
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                </div>
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                <b>Figure 3: </b> <b> Flow cytometry results of the 10 mutant promoters with the highest predictive strength. </b>
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            </div>
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        </div>
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    </div>
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<br>
  
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h2Conclusionh2
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</p>
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pAccording to fluorescence assay result, it can be found that the improvement of promoter PnisA by our software tool is a great success, though the effectiveness of PnisRRH07 and PnisRRH10 is relatively low. This helps us a lot in providing an improve to the composite part J23100-nisK-nisR+PnisA-EGFP(BBa_K4307013) and create better nisin induced expression system in iE.colii.p
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<h2>Conclusion</h2>
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<p>According to fluorescence assay result, it can be found that the improvement of promoter PnisA by our software tool is a great success, though the effectiveness of PnisRRH07 and PnisRRH10 is relatively low. This helps us a lot in providing an improve to the composite part J23100-nisK-nisR+PnisA-EGFP(BBa_K4307013) and create better nisin induced expression system in <i>E.coli</i>.</p>
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</html>
  
  

Revision as of 11:42, 13 October 2022


PnisRRH07

This part is derived from PnisA, which is originated from Lactococcus lactis subsp. Lactis, by the improvement done by our own software tool to enhance the promoter.


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]



Characterization

The following figure demonstrates our successful construction.


Figure 1: The construction results of PnisRRH07.

Fluorescence assay was done to characterize the bio brick.

We introduced these 10 promoters into our system into MACH1-T1 strain to detect EGFP fluorescence signal with and without nisin induction by microplate reader. Results demonstrated that fluorescence signals were enhanced both with and without nisin induction except for PnisRRH07 and PnisRRH10(Figure 2).


Figure 2: Fluorescence spectrophotometry results of the 10 mutant promoters with the highest predictive strength.

Furthermore, flow cytometry results confirmed that percentages of positive fluorescence signal bacteria also increased both with and without nisin induction(Figure 3), indicating that the improvement of promoter PnisA by our software tool was practical.
Figure 3: Flow cytometry results of the 10 mutant promoters with the highest predictive strength.

Conclusion

According to fluorescence assay result, it can be found that the improvement of promoter PnisA by our software tool is a great success, though the effectiveness of PnisRRH07 and PnisRRH10 is relatively low. This helps us a lot in providing an improve to the composite part J23100-nisK-nisR+PnisA-EGFP(BBa_K4307013) and create better nisin induced expression system in E.coli.