Difference between revisions of "Part:BBa K4156127"

 
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HlpA is a binding protein from Streptococcus gallolyticus that binds to acetyl heparan sulfate glycoprotein (HSPG) on the tumor surface (specifically to syndecan 1, which is upregulated during carcinogenesis). HlpA has been shown to enhance the ability of engineered bacteria to penetrate colorectal tumor cells.We added the red fluorescent protein after the HlpA gene for subsequent visualization of bacterial adhesion experiments.
 
HlpA is a binding protein from Streptococcus gallolyticus that binds to acetyl heparan sulfate glycoprotein (HSPG) on the tumor surface (specifically to syndecan 1, which is upregulated during carcinogenesis). HlpA has been shown to enhance the ability of engineered bacteria to penetrate colorectal tumor cells.We added the red fluorescent protein after the HlpA gene for subsequent visualization of bacterial adhesion experiments.
  
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===Usage and Biology===
 
===Usage and Biology===
  
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===Characterization===
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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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<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.png" 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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<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.png" 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.png" 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>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4156127 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4156127 SequenceAndFeatures</partinfo>

Revision as of 06:50, 11 October 2022


HlpA with INP-mRFP

HlpA is a binding protein from Streptococcus gallolyticus that binds to acetyl heparan sulfate glycoprotein (HSPG) on the tumor surface (specifically to syndecan 1, which is upregulated during carcinogenesis). HlpA has been shown to enhance the ability of engineered bacteria to penetrate colorectal tumor cells.We added the red fluorescent protein after the HlpA gene for subsequent visualization of bacterial adhesion experiments.

Usage and Biology

Characterization

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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 452
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 194
    Illegal NgoMIV site found at 276
    Illegal NgoMIV site found at 527
    Illegal AgeI site found at 855
    Illegal AgeI site found at 1508
    Illegal AgeI site found at 1620
  • 1000
    COMPATIBLE WITH RFC[1000]