Difference between revisions of "Part:BBa K5143007"
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ScCBD_cex'V1 : a Cellulose Binding Domain optimised for <i>Saccharomyces cerevisiae</i>, that enables the fixation of proteins to Bacterial Cellulose
 
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     <h1>Description</h1>
 
     <h1>Description</h1>
 
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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>.
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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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             <img src="https://static.igem.wiki/teams/5143/bba-k5143016-bt-toxin-cbd.png" width="200" alt="Plu1537">
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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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             <figcaption>Figure 1: Production of the fused plu1537</figcaption>
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             <figcaption>Figure 1: ScCBD_Cex'V1 three-dimensional structure</figcaption>
 
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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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            <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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     <h1>Construction</h1>
 
     <h1>Construction</h1>
 
     <p>
 
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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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       ScCBD-Cex'V1 was synthesised and its nucleotide sequence optimised for synthesis and expression in <i>Saccharomyces cerevisiae</i>. ScCDB-Cex'V1 protein is used in fusion with bioglue composite biobrick (Cp19k_MaSp1) in order to functionalise bacterial cellulose with sticky properties: <a href="https://parts.igem.org/Part:BBa_K5143022" target="_blank">BBa_K5143022</a>
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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>.
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     <h1>References</h1>
 
     <h1>References</h1>
 
     <p>
 
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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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        1. Ong, E., Gilkes, N. R., Miller, R. C., Warren, R. A. & Kilburn, D. G. Te
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cellulose-binding domain (CBD(Cex)) of an exoglucanase from Cellulomonas
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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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fmi: production in Escherichia coli and characterization of the polypeptide.  
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Biotechnol. Bioeng. 42, 401–409 (1993). <br>
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3. M. S. Kelker et al., PLOS ONE 9, (2014).
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<br>
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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.
 
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<h1>Sequence and Features</h1>
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<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K5143007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5143007 SequenceAndFeatures</partinfo>
  

Revision as of 16:51, 27 September 2024

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


Protein Description ScCBD_cex'V1 : a Cellulose Binding Domain optimised for Saccharomyces cerevisiae, that enables the fixation of proteins to Bacterial Cellulose

Description

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.

CBD_CexV1
Figure 1: ScCBD_Cex'V1 three-dimensional structure
Functional cellulose
Figure 2: Functional cellulose
ScCBD_Cex coding sequence
Figure 3: ScCBD_Cex coding sequence

Construction

ScCBD-Cex'V1 was synthesised and its nucleotide sequence optimised for synthesis and expression in Saccharomyces cerevisiae. ScCDB-Cex'V1 protein is used in fusion with bioglue composite biobrick (Cp19k_MaSp1) in order to functionalise bacterial cellulose with sticky properties: BBa_K5143022

References

1. Ong, E., Gilkes, N. R., Miller, R. C., Warren, R. A. & Kilburn, D. G. Te cellulose-binding domain (CBD(Cex)) of an exoglucanase from Cellulomonas fmi: production in Escherichia coli and characterization of the polypeptide. Biotechnol. Bioeng. 42, 401–409 (1993).

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.


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]