Difference between revisions of "Part:BBa K4156113"
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In this biobrick, we used the plac operon to regulate downstream mRFP expression and eventually introduced the rrnB T1 and T7Te dual terminators. this design allowed the spatiotemporal intensity of plac manipulator-regulated gene expression to be measured by the red fluorescent signal. We were thus able to verify the specific response of the plac operon to lactose.
 
In this biobrick, we used the plac operon to regulate downstream mRFP expression and eventually introduced the rrnB T1 and T7Te dual terminators. this design allowed the spatiotemporal intensity of plac manipulator-regulated gene expression to be measured by the red fluorescent signal. We were thus able to verify the specific response of the plac operon to lactose.
  
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===Characterization===
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We first successfully isolated this part and verified it by gel electrophoresis before fusing mRFP with plac.
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<html>
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/rfp.jpg" alt="control">
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                <figcaption><b>Figure 1:</b> Agarose gel electrophoresis result of mRFP.</figcaption>
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              </figure>
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</html>
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==Enhancement of E. coli EcN tropism for colorectal cancer==
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To make the engineered strain nengg better adhere to colorectal cancer cells, we used engineered bacteria carrying the adhesion proteins INP-HlpA and mRFP, and the fluorescence was significantly enhanced compared to the control engineered bacteria expressing mRFP alone, indicating that the HlpA protein could effectively improve the adhesion of EcN to colorectal cells. (where CT26 and RKO were magnified 400-fold, while RKO was magnified 300-fold), and this change was particularly evident in RKO cells.
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<html>
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/13-1.jpg" alt="control">
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                <figcaption><b>Figure 1 :</b> Fluorescence intensity of engineered bacteria carrying INP-HlpA and mRFP, versus control engineered bacteria expressing mRFP (CT26 were magnified 400-fold) </figcaption>
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              </figure>
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</html>
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<html>
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/13-2.jpg" alt="control">
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                <figcaption><b>Figure 2:</b> Fluorescence intensity of engineered bacteria carrying INP-HlpA and mRFP, versus control engineered bacteria expressing mRFP (RKO were magnified 300-fold)</figcaption>
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              </figure>
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</html>
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<html>
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/13-3.jpg" alt="control">
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                <figcaption><b>Figure 3:</b>Fluorescence intensity of engineered bacteria carrying INP-HlpA and mRFP, versus control engineered bacteria expressing mRFP (SW480 were magnified 400-fold)</figcaption>
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              </figure>
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</html>
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 02:51, 12 October 2022


plac-mRFP

plac-mRFP is a biobrick that expresses mRFP controlled by the plac operon ( BBa_R0010 ). plac-mRFP functions as a red fluorescent signal to characterize the performance of the plac operon


Usage and Biology

In this biobrick, we used the plac operon to regulate downstream mRFP expression and eventually introduced the rrnB T1 and T7Te dual terminators. this design allowed the spatiotemporal intensity of plac manipulator-regulated gene expression to be measured by the red fluorescent signal. We were thus able to verify the specific response of the plac operon to lactose.


Characterization

We first successfully isolated this part and verified it by gel electrophoresis before fusing mRFP with plac.

control
Figure 1: Agarose gel electrophoresis result of mRFP.

Enhancement of E. coli EcN tropism for colorectal cancer

To make the engineered strain nengg better adhere to colorectal cancer cells, we used engineered bacteria carrying the adhesion proteins INP-HlpA and mRFP, and the fluorescence was significantly enhanced compared to the control engineered bacteria expressing mRFP alone, indicating that the HlpA protein could effectively improve the adhesion of EcN to colorectal cells. (where CT26 and RKO were magnified 400-fold, while RKO was magnified 300-fold), and this change was particularly evident in RKO cells.


control
Figure 1 : Fluorescence intensity of engineered bacteria carrying INP-HlpA and mRFP, versus control engineered bacteria expressing mRFP (CT26 were magnified 400-fold)


control
Figure 2: Fluorescence intensity of engineered bacteria carrying INP-HlpA and mRFP, versus control engineered bacteria expressing mRFP (RKO were magnified 300-fold)


control
Figure 3:Fluorescence intensity of engineered bacteria carrying INP-HlpA and mRFP, versus control engineered bacteria expressing mRFP (SW480 were magnified 400-fold)

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 864
    Illegal AgeI site found at 976
  • 1000
    COMPATIBLE WITH RFC[1000]