Difference between revisions of "Part:BBa K5117022"
 
(5 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K5117022 short</partinfo>
 
<partinfo>BBa_K5117022 short</partinfo>
  
−
This parts contains the <i>cotY</i> gene of <i>Bacillus subtilis</i> followed by a 10 bp spacer.
+
This parts contains the <i>cotY</i> gene out of the <i>cotYZ</i> gene cluster of <i>Bacillus subtilis</i>.
−
 
+
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.  
−
The <i>cotY</i> gene is located in the <i>cotYZ</i> gene cluster of <i>B. subtilis</i>.
+
Information regarding the <i>cotYZ</i> operon can be found in the following databases:
−
 
+
−
Information regarding the <i>cotYZ</i> operon can be found on the following databases:
+
 
<html><a href="https://subtiwiki.uni-goettingen.de/">SubtiWiki</a></html>,  
 
<html><a href="https://subtiwiki.uni-goettingen.de/">SubtiWiki</a></html>,  
 
<html><a href="https://dbtbs.hgc.jp/">DBTBS</a></html>.
 
<html><a href="https://dbtbs.hgc.jp/">DBTBS</a></html>.
  
−
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).
 
  
−
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).  
+
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).  
 +
 
  
 
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).  
 
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).  
Line 39: Line 37:
 
<partinfo>BBa_K5117022 parameters</partinfo>
 
<partinfo>BBa_K5117022 parameters</partinfo>
 
<!-- -->
 
<!-- -->
−
 
−
===References===
 

Latest revision as of 08:45, 30 September 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).


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