Difference between revisions of "Part:BBa K5143007"
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     <h1>Description</h1>
 
     <h1>Description</h1>
 
     <p>
 
     <p>
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         ScCBD_cex'V1 (Cellulose Binding Domain) is a peptide that has a huge affinity for bacterial cellulose. When fused with another protein, ScCBD_cex'V1 enables the fixation of the protein on the bacterial cellulose.
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         The final part pUC57-5'URA-GAP_promoter-alphafactorV1-plu1537-dCBD-TDH1_term-3'URA is composed of the plu1537 gene is a antitermite toxin (known as Bt toxin) fused to a dCBD and an alphafactor signal. <br> This final construct has been transformed in the BY4741 <i> S. cerevisiae </i> strain. This transformed yeast will help us to functionalize bacterial cellulose with antitermite properties by a coculture with the yeast and the cellulose producing bacteria <i> Komagataeibacter rhaeticus </i>.
 
     </p>
 
     </p>
 
     <div class="image-container">
 
     <div class="image-container">
 
         <figure>
 
         <figure>
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             <img src="https://static.igem.wiki/teams/5143/bba-k5143007-cbd-cexv1-1.png" width="200" alt="CBD_CexV1">
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             <img src="https://static.igem.wiki/teams/5143/bba-k5143016-bt-toxin-cbd.png" width="200" alt="Plu1537">
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             <figcaption>Figure 1: ScCBD_Cex'V1 three-dimensional structure</figcaption>
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             <figcaption>Figure 1: Production of the fused plu1537</figcaption>
 
         </figure>
 
         </figure>
 
         <figure>
 
         <figure>
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             <img src="https://static.igem.wiki/teams/5143/bba-k5143007-cbd-cexv1-2.png" width="400" alt="Functional cellulose">
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            <figcaption>Figure 2: Functional cellulose</figcaption>
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        </figure>
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        <figure>
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            <img src="https://static.igem.wiki/teams/5143/bba-k5143007-cbd-cexv1-3.png" width="100" alt="ScCBD_Cex coding sequence">
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            <figcaption>Figure 3: ScCBD_Cex coding sequence</figcaption>
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</figure>
 
</figure>
  
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     <h1>Construction</h1>
 
     <h1>Construction</h1>
 
     <p>
 
     <p>
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       ScCBD-Cex'V1 was synthesised and its nucleotide sequence optimised for synthesis and expression in <i>Saccharomyces cerevisiae</i>. This protein is used in fusion with a yellow chromoprotein (fwYellow) in order to color the bacterial cellulose in yellow: <a href="https://parts.igem.org/Part:BBa_K5143023" target="_blank">BBa_K5143023</a>
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       pUC57-5'URA-GAP_promoter-alphafactorV1-plu1537-dCBD-TDH1_term-3'URA is composed of the pUC57 backbone  <a href="https://parts.igem.org/Part:BBa_K5143005" target="_blank">BBa_K5143005</a> and the alphafactor-plu1537-dCBD composite part <a href="https://parts.igem.org/Part:BBa_K5143016" target="_blank">BBa_K5143016</a>. <br>
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The alphafactor-plu1537-dCBD composite part was synthesised and its nucleotide sequence optimised for synthesis and expression in <i>Saccharomyces cerevisiae</i>.
 
     </p>
 
     </p>
 
     <h1>References</h1>
 
     <h1>References</h1>
 
     <p>
 
     <p>
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        Ong, E., Gilkes, N. R., Miller, R. C., Warren, R. A. & Kilburn, D. G. Te
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      1. Linder, M.; Salovuori, I.; Ruohonen, L.; Teeri, T.T., 1996. Characterization of a Double Cellulose-binding Domain. SYNERGISTIC HIGH AFFINITY BINDING TO CRYSTALLINE CELLULOSE. Journal of Biological Chemistry, 271(35), pp.21268–21272. Available at: http://www.jbc.org/content/271/35/21268.full <br>
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cellulose-binding domain (CBD(Cex)) of an exoglucanase from Cellulomonas
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fmi: production in Escherichia coli and characterization of the polypeptide.  
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2. Gilbert, C.; Tang, T.-C.; Ott, W.; Dorr, B. A.; Shaw, W. M.; Sun, G. L.; Lu, T. K.; Ellis, T. Living Materials with Programmable Functionalities Grown from Engineered Microbial Co-Cultures. Nat. Mater. 2021, 20 (5), 691–700. https://doi.org/10.1038/s41563-020-00857-5. <br>
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Biotechnol. Bioeng. 42, 401–409 (1993).  
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Gilbert, C.; Tang, T.-C.; Ott, W.; Dorr, B. A.; Shaw, W. M.; Sun, G. L.; Lu, T. K.; Ellis, T. Living Materials with Programmable Functionalities Grown from Engineered Microbial Co-Cultures. Nat. Mater. 2021, 20 (5), 691–700. https://doi.org/10.1038/s41563-020-00857-5.
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3. M. S. Kelker et al., PLOS ONE 9, (2014).
 
     </p>
 
     </p>
 
</body>
 
</body>

Revision as of 16:37, 27 September 2024

ScCBD_cex'V1 : a Cellulose Binding Domain that enables the fixation of proteins to Cellulose


Protein Description

Description

The final part pUC57-5'URA-GAP_promoter-alphafactorV1-plu1537-dCBD-TDH1_term-3'URA is composed of the plu1537 gene is a antitermite toxin (known as Bt toxin) fused to a dCBD and an alphafactor signal.
This final construct has been transformed in the BY4741 S. cerevisiae strain. This transformed yeast will help us to functionalize bacterial cellulose with antitermite properties by a coculture with the yeast and the cellulose producing bacteria Komagataeibacter rhaeticus .

Plu1537
Figure 1: Production of the fused plu1537

Construction

pUC57-5'URA-GAP_promoter-alphafactorV1-plu1537-dCBD-TDH1_term-3'URA is composed of the pUC57 backbone BBa_K5143005 and the alphafactor-plu1537-dCBD composite part BBa_K5143016.
The alphafactor-plu1537-dCBD composite part was synthesised and its nucleotide sequence optimised for synthesis and expression in Saccharomyces cerevisiae.

References

1. Linder, M.; Salovuori, I.; Ruohonen, L.; Teeri, T.T., 1996. Characterization of a Double Cellulose-binding Domain. SYNERGISTIC HIGH AFFINITY BINDING TO CRYSTALLINE CELLULOSE. Journal of Biological Chemistry, 271(35), pp.21268–21272. Available at: http://www.jbc.org/content/271/35/21268.full
2. Gilbert, C.; Tang, T.-C.; Ott, W.; Dorr, B. A.; Shaw, W. M.; Sun, G. L.; Lu, T. K.; Ellis, T. Living Materials with Programmable Functionalities Grown from Engineered Microbial Co-Cultures. Nat. Mater. 2021, 20 (5), 691–700. https://doi.org/10.1038/s41563-020-00857-5.
3. M. S. Kelker et al., PLOS ONE 9, (2014).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 256
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 226
    Illegal AgeI site found at 265
  • 1000
    COMPATIBLE WITH RFC[1000]