Part:BBa_K5117022

CotY

This parts contains the cotY gene out of the cotYZ gene cluster of Bacillus subtilis. The coding sequence (CDS) of cotY is followed by a spacer consisting of 10 bp of the natural genome sequence downstream from the cotYZ operon. This creates space before the terminator and ensures that the ribosome is able to read the full length of the CDS.

Information regarding the cotYZ operon can be found on the following databases: SubtiWiki, DBTBS.

The encoded protein CotY is a spore coat protein of B. subtilis, which is able to form endospores. CotY is located in the crust, the outermost spore layer, and has been shown to be well suited for protein immobilization (McKenney et al. 2013, Bartels et al. 2018, Lin et al. 2020). This part has been designed for the construction of fusion proteins, expecially for spore surface diplay. As this part contains a stop codon, it is only suited for N-terminal fusions. N-terminal fusions provided higher reporter signals than C-terminal fusions when working with sfGFP (Bartels et al. 2018).

CotY only served for design purposes of the TU Dresden iGEM 2024 Team and was required for the construction of composite parts (see Contribution page).

Target organism: Bacillus subtilis

Main purpose of use:: Gene expression and production of fusion proteins (especially for spore surface display)

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 23
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

Bartels J., López Castellanos S., Radeck J., Mascher T. (2018): Sporobeads: The utilization of the Bacillus subtilis endospore crust as a protein display platform. ACS synthetic biology 7(2), 452-461. https://doi.org/10.1021/acssynbio.7b00285

McKenney P. T., Driks A., Eichenberger P. (2013): The Bacillus subtilis endospore: assembly and functions of the multilayered coat. Nature Reviews Microbiology 11 (1), 33–44. https://doi.org/10.1038/nrmicro2921

Lin P., Yuan H., Du J., Liu K., Liu H., Wang T. (2020): Progress in research and application development of surface display technology using Bacillus subtilis spores. Applied microbiology and biotechnology 104 (6), 2319–2331. https://doi.org/10.1007/s00253-020-10348-x