Difference between revisions of "Part:BBa K5269023"
 
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<partinfo>BBa_K5269023 short</partinfo>
 
<partinfo>BBa_K5269023 short</partinfo>
  
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It contains SD sequences(BBa_K5269034), SpyTag (BBa_K5269017), Mgfp-5 (BBa_5269013) and 6xHis sequences. In addition, the BsmB I cleavage sites at both ends of the sequence allow for Golden Gate Assembly to the Golden Gate Assembly Site (BBa_K5269020) and T7 promoter-shRNA-T7 terminator-Golden Gate Assembly Site(BBa_K5269025). The SpyTag/SpyCatcher system works when used with Lpp-ompA-SpyCatcher(BBa_K5269026) or INPNC-SpyCatcher(BBa_K5269027). Therefore, we expect the adhesion protein Mgfp-5 to be stable on the membrane vesicles, which will facilitate the retention of our shRNA-encapsulated membrane vesicles on the plant surface.
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It contains SD-sequences (BBa_K5269034), SpyTag, Mgfp-5 (BBa_K5269013), and His-Tag sequences. In addition, the BsmB I cleavage sites at both ends of the sequence allow for Golden Gate Assembly to the Golden Gate Assembly Site (BBa_K5269020) and T7 promoter-shRNA-T7 terminator-Golden Gate Assembly Site (BBa_K5269025). The SpyCatcher-SpyTag system works when used with Lpp-OmpA-SpyCatcher (BBa_K5269026) or INPNC-SpyCatcher (BBa_K5269027). Therefore, we expect the adhesion protein Mgfp-5 to be stable on the Membrane Vesicles, which will facilitate the retention of our shRNA-encapsulated membrane vesicles on the plant surface.
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Here are the results of SDS-PAGE of OmpA-Mgfp5 and INP-Mgfp5 bound by SpyCatcher-SpyTag. See our results page (https://2024.igem.wiki/tokyotech/results/) for details.
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<table border="1">
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  <thead>
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    <tr>
 +
      <th>Serial</th>
 +
      <th>Stock Name</th>
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      <th>Scaffolding Protein - Display Protein</th>
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      <th>Molecular Weight (kDa)</th>
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      <th>Corresponding SDS-PAGE Lanes (Periplasmic/Cytoplasmic/Membrane Fractions)</th>
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      <th>Presence of Bands (Periplasmic/Cytoplasmic/Membrane Fractions)</th>
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    </tr>
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  </thead>
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  <tbody>
 +
    <tr>
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      <td>O_1</td>
 +
      <td>O_Mgfp5</td>
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      <td>OmpA-Mgfp5</td>
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      <td>36.8</td>
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      <td>1/2/3</td>
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      <td>Yes/Yes/Yes</td>
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    </tr>
 +
    <tr>
 +
      <td>I_1</td>
 +
      <td>I_Mgfp5</td>
 +
      <td>INP-Mgfp5</td>
 +
      <td>54.2</td>
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      <td>4/5/6</td>
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      <td>Yes/Yes/Yes</td>
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    </tr>
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    <tr>
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      <td>O_2</td>
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      <td>O_Glucanases</td>
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      <td>OmpA-Glucanases</td>
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      <td>78.5</td>
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      <td>7/8/9</td>
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      <td>Yes/Yes/Yes</td>
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    </tr>
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    <tr>
 +
      <td>I_2</td>
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      <td>I_Glucanases</td>
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      <td>INP-Glucanases</td>
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      <td>95.9</td>
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      <td>10/11/12</td>
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      <td>Yes/Yes/Yes</td>
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    </tr>
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    <tr>
 +
      <td>O_3</td>
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      <td>O_Mgfp5-Glucanases</td>
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      <td>OmpA-Glucanases, OmpA-Mgfp5</td>
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      <td>78.5, 36.8</td>
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      <td>13/14/15</td>
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      <td>No/Yes/Yes, Yes/Yes/Yes</td>
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    </tr>
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    <tr>
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      <td>I_3</td>
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      <td>I_Mgfp5-Glucanases</td>
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      <td>INP-Glucanases, INP-Mgfp5</td>
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      <td>95.9, 54.2</td>
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      <td>16/17/18</td>
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      <td>No/No/No, No/Yes/Yes</td>
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    </tr>
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    <tr>
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      <td>O_4</td>
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      <td>O_Glucanases-GH19Chitinase</td>
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      <td>OmpA-Glucanases, OmpA-GH19Chitinase</td>
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      <td>78.5, 56.5</td>
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      <td>19/20/21</td>
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      <td>No/Yes/Yes, No/No/No</td>
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    </tr>
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    <tr>
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      <td>I_4</td>
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      <td>I_Glucanases-GH19Chitinase</td>
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      <td>INP-Glucanases, INP-GH19Chitinase</td>
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      <td>95.9, 73.9</td>
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      <td>22/23/24</td>
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      <td>Yes/Yes/No, No/No/No</td>
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    </tr>
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  </tbody>
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</table>
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<html>
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<img src="https://static.igem.wiki/teams/5269/parts-registry/sdspage.webp" alt="SDS-PAGE Image" style="width:80%;>
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<br>
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</html>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 12:28, 2 October 2024


SpyTag-Mgfp-5-His-Tag and BsmB I cut site(for GGA)

It contains SD-sequences (BBa_K5269034), SpyTag, Mgfp-5 (BBa_K5269013), and His-Tag sequences. In addition, the BsmB I cleavage sites at both ends of the sequence allow for Golden Gate Assembly to the Golden Gate Assembly Site (BBa_K5269020) and T7 promoter-shRNA-T7 terminator-Golden Gate Assembly Site (BBa_K5269025). The SpyCatcher-SpyTag system works when used with Lpp-OmpA-SpyCatcher (BBa_K5269026) or INPNC-SpyCatcher (BBa_K5269027). Therefore, we expect the adhesion protein Mgfp-5 to be stable on the Membrane Vesicles, which will facilitate the retention of our shRNA-encapsulated membrane vesicles on the plant surface.

Here are the results of SDS-PAGE of OmpA-Mgfp5 and INP-Mgfp5 bound by SpyCatcher-SpyTag. See our results page (https://2024.igem.wiki/tokyotech/results/) for details.

<thead> </thead> <tbody> </tbody>
Serial Stock Name Scaffolding Protein - Display Protein Molecular Weight (kDa) Corresponding SDS-PAGE Lanes (Periplasmic/Cytoplasmic/Membrane Fractions) Presence of Bands (Periplasmic/Cytoplasmic/Membrane Fractions)
O_1 O_Mgfp5 OmpA-Mgfp5 36.8 1/2/3 Yes/Yes/Yes
I_1 I_Mgfp5 INP-Mgfp5 54.2 4/5/6 Yes/Yes/Yes
O_2 O_Glucanases OmpA-Glucanases 78.5 7/8/9 Yes/Yes/Yes
I_2 I_Glucanases INP-Glucanases 95.9 10/11/12 Yes/Yes/Yes
O_3 O_Mgfp5-Glucanases OmpA-Glucanases, OmpA-Mgfp5 78.5, 36.8 13/14/15 No/Yes/Yes, Yes/Yes/Yes
I_3 I_Mgfp5-Glucanases INP-Glucanases, INP-Mgfp5 95.9, 54.2 16/17/18 No/No/No, No/Yes/Yes
O_4 O_Glucanases-GH19Chitinase OmpA-Glucanases, OmpA-GH19Chitinase 78.5, 56.5 19/20/21 No/Yes/Yes, No/No/No
I_4 I_Glucanases-GH19Chitinase INP-Glucanases, INP-GH19Chitinase 95.9, 73.9 22/23/24 Yes/Yes/No, No/No/No


SDS-PAGE ImageSequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 381
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]