Part:BBa_K4593019:Design
Endolysin combinative expression circuit in E.coli
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2537
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2195
Illegal NgoMIV site found at 3021
Illegal NgoMIV site found at 3695
Illegal AgeI site found at 2064
Illegal AgeI site found at 3165
Illegal AgeI site found at 3393
Illegal AgeI site found at 3564
Illegal AgeI site found at 3677 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part is designed as a subpart of the "S. aureus in vivo elimination apparatus for E. coli" (BBa_K4593020). It could not show observable function independently due to the lack of a release system. The promoter, RBS, and codon are adjusted to be suitable for expression in E. coli.
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