Part:BBa_K5143007

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

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 .

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]