Difference between revisions of "Part:BBa K4212050:Design"
 
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===Design Notes===
 
===Design Notes===
  
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PsspB is a promoter, which can aid the expression of gerA. GerA is one of the germinant receptors of B.subtilis, and it can respond to the presence of L-alanine and increase the germinantion rate of B.subtilis. The previous iGEM team iGEM17_SZU-China proved the superior transcription efficiency of this promoter.  tL3S2P21 is a strong promoter, which helps to increase the production of desired protein. We included the operon for germinant receptor GerA because GerA is responsive to L-alanine and responsible for B.subtilis germination (Ramírez-Guadiana et al., 2017). Synthetic operon designed with GerAA, GerKB and GerAC coding sequences from the B. subtilis genome. Designed to be transformed in a B. subtilis knockout strain so as to produce a germinant receptor that contains a binding site for glucose, and when activated triggers an orthogonal signalling cascade resulting in germination.  
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PsspB is a promoter, which can aid the expression of gerA. GerA is one of the germinant receptors of B.subtilis, and it can respond to the presence of L-alanine and increase the germinantion rate of B.subtilis. However, we planned to engineer the receptor to make it more sensitive to N-acetylglucosamine (NAG) - the chitin monomer. The previous iGEM team iGEM17_SZU-China proved the superior transcription efficiency of this promoter.  tL3S2P21 is a strong terminator, which helps to increase the production of desired protein. We included the operon for germinant receptor GerA because GerA is responsive to L-alanine and responsible for B.subtilis germination (Ramírez-Guadiana et al., 2017). Synthetic operon designed with GerAA, GerKB and GerAC coding sequences from the B. subtilis genome. Designed to be transformed in a B. subtilis knockout strain so as to produce a germinant receptor that contains a binding site for glucose, and when activated triggers an orthogonal signalling cascade resulting in germination.  
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Latest revision as of 11:24, 9 October 2022


Ger2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 934
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 934
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 934
    Illegal BamHI site found at 919
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 934
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 934
    Illegal NgoMIV site found at 66
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

PsspB is a promoter, which can aid the expression of gerA. GerA is one of the germinant receptors of B.subtilis, and it can respond to the presence of L-alanine and increase the germinantion rate of B.subtilis. However, we planned to engineer the receptor to make it more sensitive to N-acetylglucosamine (NAG) - the chitin monomer. The previous iGEM team iGEM17_SZU-China proved the superior transcription efficiency of this promoter. tL3S2P21 is a strong terminator, which helps to increase the production of desired protein. We included the operon for germinant receptor GerA because GerA is responsive to L-alanine and responsible for B.subtilis germination (Ramírez-Guadiana et al., 2017). Synthetic operon designed with GerAA, GerKB and GerAC coding sequences from the B. subtilis genome. Designed to be transformed in a B. subtilis knockout strain so as to produce a germinant receptor that contains a binding site for glucose, and when activated triggers an orthogonal signalling cascade resulting in germination.


Source

Synthetic construct.

References

[1] https://parts.igem.org/Part:BBa_K2232020 [2] Ramírez-Guadiana, F.H., Meeske, A.J., Wang, X., Rodrigues, C.D.A. & Rudner, D.Z. (2017) The Bacillus subtilis germinant receptor GerA triggers premature germination in response to morphological defects during sporulation. Molecular Microbiology. 105 (5), 689–704. doi:10.1111/mmi.13728.