Part:BBa_K5322038
pET29a-LuxI-PhiX174E-pDawn-MazF
Usage and Biology
In order to achieve the best effect of bacterial treatment, we designed the PhiX174E lysis system pLuxⅠ-RBS-PhiX174E-rrnB T1 regulated by pLuxI. When the engineered bacteria detect NO, the mussel foot protein Mfp is synthesized, so that the probiotic EcN is fixed to the site of enteritis, thus forming a dominant bacterial community. However, the therapeutic protein SOD cannot cross the membrane. To ensure its performance, we use the principle of bacterial group response, so that EcN in the site of enteritis increases due to the increase in population density, activates the group response system, expresses PhiX174E protein, and then promotes bacterial lysis to release therapeutic proteins, achieving the best therapeutic effect.
Biosafety has always been a focus of biologists. In order to protect biosafety and avoid potential problems such as genetic contamination, we designed the pDawn-MzaF safety module. pDawn is a blue light-inducible promoter that can activate the expression of downstream genes under the irradiation of natural light or a single blue light source. MazF is an RNA enzyme that can efficiently decompose bacterial mRNA, thereby inhibiting bacterial growth and reproduction and other life activities.
To this end, we designed the plasmid pET29a-LuxI-PhiX174E-pDawn-MazF, as shown in the figure 1-1, to achieve the effects of lysis and suicide. We successfully constructed the plasmid, and at the same time, to explore the performance of the group response system, we measured the group response growth curve to further prove the feasibility of the project,as shown in the figure 1-2.
Figure 1-1 Plasmid pET29a-LuxI-PhiX174E-pDawn-MazF
Figure 1-2 Population response CFU results
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 826
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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