Difference between revisions of "Part:BBa K5143028"
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     <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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      Our team has nicknamed this plasmid “<b>Bt toxin plasmid</b>”. 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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Revision as of 16:55, 27 September 2024


-- No description --


Protein Description

Description

Our team has nicknamed this plasmid “Bt toxin plasmid”. 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
    INCOMPATIBLE WITH RFC[10]
    Illegal suffix found in sequence at 1927
    Illegal EcoRI site found at 5366
    Illegal EcoRI site found at 5893
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5366
    Illegal EcoRI site found at 5893
    Illegal SpeI site found at 1928
    Illegal PstI site found at 1942
    Illegal NotI site found at 1935
    Illegal NotI site found at 5899
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5366
    Illegal EcoRI site found at 5893
    Illegal XhoI site found at 3531
    Illegal XhoI site found at 5372
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal suffix found in sequence at 1928
    Illegal EcoRI site found at 5366
    Illegal EcoRI site found at 5893
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 5893
    Illegal EcoRI site found at 5366
    Illegal SpeI site found at 1928
    Illegal PstI site found at 1942
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2640