Difference between revisions of "Part:BBa K4593018:Design"

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===References===
 
===References===
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[1] Chang, Y., Li, Q., Zhang, S., Zhang, Q., Liu, Y., Qi, Q., & Lu, X. (2023). Identification and Molecular Modification of Staphylococcus aureus Bacteriophage Lysin LysDZ25. ACS Infectious Diseases, 9(3), 497–506. https://doi.org/10.1021/acsinfecdis.2c00493
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[2] Gu, J., Feng, Y., Feng, X., Sun, C., Lei, L., Ding, W., Niu, F., Jiao, L., Yang, M., Li, Y., Liu, X., Song, J., Cui, Z., Dong Soo Han, Du, C., Yang, Y., Liu, Z.-J., Liu, Z.-J., & Han, W. (2014). Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented “EF-Hand-Like” Calcium-Binding Phage Lysin. 10(5), e1004109–e1004109. https://doi.org/10.1371/journal.ppat.1004109
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[3] Lee, Chanyoung, et al. “Development of Advanced Chimeric Endolysin to Control Multidrug-Resistant Staphylococcus Aureus through Domain Shuffling.” ACS Infectious Diseases, vol. 7, no. 8, 28 May 2021, pp. 2081–2092. https://doi.org/10.1021/acsinfecdis.0c00812

Latest revision as of 23:45, 8 October 2023


Endolysin combinative expression circuit in B. subtilis


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2576
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2228
    Illegal NgoMIV site found at 3060
    Illegal NgoMIV site found at 3734
    Illegal AgeI site found at 2097
    Illegal AgeI site found at 3204
    Illegal AgeI site found at 3432
    Illegal AgeI site found at 3603
    Illegal AgeI site found at 3716
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is designed as a subpart of the "S. aureus in vivo elimination apparatus for B. subtilis" (BBa_K4593021). It could not show observable function independently due to the lack of a release system. The promoter and RBS are adjusted to be suitable for expression in B. subtilis. However, further codon optimization might be needed to achieve full expression potential.


Source

LysDZ25 is from bacteriophage DZ25[1], LysGH15 is from Bacteriophage GH15[2], and ClyC is a hybrid endolysin with CBD of LysPALS1 and EAD of LysSA12[3].

References

[1] Chang, Y., Li, Q., Zhang, S., Zhang, Q., Liu, Y., Qi, Q., & Lu, X. (2023). Identification and Molecular Modification of Staphylococcus aureus Bacteriophage Lysin LysDZ25. ACS Infectious Diseases, 9(3), 497–506. https://doi.org/10.1021/acsinfecdis.2c00493

[2] Gu, J., Feng, Y., Feng, X., Sun, C., Lei, L., Ding, W., Niu, F., Jiao, L., Yang, M., Li, Y., Liu, X., Song, J., Cui, Z., Dong Soo Han, Du, C., Yang, Y., Liu, Z.-J., Liu, Z.-J., & Han, W. (2014). Structural and Biochemical Characterization Reveals LysGH15 as an Unprecedented “EF-Hand-Like” Calcium-Binding Phage Lysin. 10(5), e1004109–e1004109. https://doi.org/10.1371/journal.ppat.1004109

[3] Lee, Chanyoung, et al. “Development of Advanced Chimeric Endolysin to Control Multidrug-Resistant Staphylococcus Aureus through Domain Shuffling.” ACS Infectious Diseases, vol. 7, no. 8, 28 May 2021, pp. 2081–2092. https://doi.org/10.1021/acsinfecdis.0c00812