Difference between revisions of "Part:BBa K5117022"
 
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<partinfo>BBa_K5117022 short</partinfo>
 
<partinfo>BBa_K5117022 short</partinfo>
  
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The protein CotY originated from <i>Bacillus subtilis</i>, which is able to form endospores.
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This parts contains the <i>cotY</i> gene out of the <i>cotYZ</i> gene cluster of <i>Bacillus subtilis</i>.
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CotY is a spore coat protein located in the outermost layer, the spore crust.
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The coding sequence (CDS) of <i>cotY</i> is followed by a spacer consisting of 10 bp of the natural genome sequence downstream from the <i>cotYZ</i> operon. This creates space before the terminator and ensures that the ribosome is able to read the full length of the CDS.
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Its coding sequence is part of the <i>cotYZ</i> operon in the <i>B. subtilis</i> genome.
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Information regarding the <i>cotYZ</i> operon can be found in the following databases:
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<html><a href="https://subtiwiki.uni-goettingen.de/">SubtiWiki</a></html>,
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<html><a href="https://dbtbs.hgc.jp/">DBTBS</a></html>.
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The encoded protein CotY is a spore coat protein of <i>B. subtilis</i>, 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 <i>et al.</i> 2013, Bartels <i>et al.</i> 2018, Lin <i>et al.</i> 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 <i>et al.</i> 2018).
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CotY only served for design purposes of the TU Dresden iGEM 2024 Team and was required for the construction of composite parts (see <html><a href="https://2024.igem.wiki/tu-dresden/contribution">Contribution</a></html> page).
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<b>Biosafety level:</b> S1
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<b>Target organism:</b> <i>Bacillus subtilis</i>
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<b>Main purpose of use:</b> Anchor protein for the construction of fusion proteins in <i>B. subtilis</i>
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<b> Potential application:</b> Spore surface display
  
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K5117022 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5117022 SequenceAndFeatures</partinfo>
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===References===
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Bartels J., López Castellanos S., Radeck J., Mascher T. (2018): Sporobeads: The utilization of the <i>Bacillus subtilis</i> endospore crust as a protein display platform. ACS synthetic biology 7(2), 452-461. https://doi.org/10.1021/acssynbio.7b00285
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McKenney P. T., Driks A., Eichenberger P. (2013): The <i>Bacillus subtilis</i> endospore: assembly and functions of the multilayered coat. Nature Reviews Microbiology 11 (1), 33–44. https://doi.org/10.1038/nrmicro2921
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Lin P., Yuan H., Du J., Liu K., Liu H., Wang T. (2020): Progress in research and application development of surface display technology using <i>Bacillus subtilis</i> spores. Applied microbiology and biotechnology 104 (6), 2319–2331. https://doi.org/10.1007/s00253-020-10348-x
  
  

Latest revision as of 23:58, 1 October 2024


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 in 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).


Biosafety level: S1

Target organism: Bacillus subtilis

Main purpose of use: Anchor protein for the construction of fusion proteins in B. subtilis

Potential application: 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