Part:BBa_K5322002
_NOTOC__ Constitutive Mfp3 Expression System
Nitric-oxide-inducible lysis module
Contents
Usage and Biology
Plasm11:14, 29 September 2024 (UTC)~
Construction of the plasmid
In o3119-``````````
Figure 2-1 Plasmid pET29(a)-pJ23119-SoxR-T-pSoxS-RBS-Mfp3-T7
Figure 2-2 Colony PCR gel electrophoresis of plasmid pET29(a)-pJ23119-SoxR-T-pSoxS-RBS-PhiX174E-T7(1320bp)
Figure 2-3 plasmid pET29(a)-pJ23119-SoxR-T-pSoxS-RBS-PhiX174E-T7 sequencing result
Protein Expression Validation
We verified the performance of the lysis module through dry experiments and plan to complete wet experimental validation of its lytic function in the future. Through literature review and numerical modeling validation, we found that after inducing the expression of antimicrobial peptides and lysis proteins with NO for twenty minutes, the engineered strain will be lysed by the lysis protein and release the antimicrobial peptides. Mathematical modeling confirmed that at this time, the concentration of antimicrobial peptides is sufficient to reach an effective inhibitory concentration.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 572 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1009
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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