Part:BBa_K5238015
Expression System and Induced Suicide Switch
This composite part combines our previous suicide switch by first expressing the target gene FolKE under the P9 promoter, and then regulating it with the suicide switch. It mainly includes the following components 1.P9 P9 is the strongest promoter of Lactobacillus plantarum, which can induce the expression of downstream genes 2.FolKE Production of tetrahydrofolate as our target product to bind to target lilrb3 for the treatment of Alzheimer's disease 3.PfdhF The low oxygen promoter PfdhF simulates the low oxygen environment in the intestine by sensing the oxygen concentration in the surrounding environment, and expresses the downstream protein SrpR under low oxygen conditions 4.SrpR The repressor protein SrpR inhibits the downstream P-srpR promoter, thereby achieving regulation 5. P-SrpR The promoter P-SrpR can be regulated by a repressor protein. If the repressor protein is expressed upstream, the promoter will not express it 6.LysKB317 The core of our suicide switch is the endosomal gene LysKB317, which is derived from the autophagic bacterial genome. Endolysin is an enzyme that degrades bacterial cell walls, leading to cell lysis. In our design, the expression of LysKB317 is strictly regulated by an inducible promoter, ensuring that cell lysis only occurs under non permissive conditions, such as a lack of essential nutrients or the presence of specific inducers.
We hope to achieve the expression of FolKE gene through this composite part to produce tetrahydrofolate binding specific receptors for Alzheimer's disease, achieving the cure of related neurological diseases, while ensuring the controllability of bacterial cell lysis through the action of suicide switches. This system enhances biosafety by preventing accidental transmission of genetically modified organisms. Our experiment validated the reliability and effectiveness of the suicide switch, making it a robust biological closure tool in synthetic biology applications.
Due to the limited research on Lactobacillus plantarum, there is even less research on its suicide switch. And we found the endosome LysVNet 17 targeting Lactobacillus plantarum, which is almost harmless to the human body, ensuring the safety of Lactobacillus plantarum in treating human diseases and avoiding biological leakage. Our suicide switch system can not only be used in our system, but also in most synthetic biology systems that use lactobacilli. This indicates that plant lactobacilli have finally found a suitable suicide switch, which has broad application and impact significance.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1048
Illegal EcoRI site found at 1191 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1048
Illegal EcoRI site found at 1191
Illegal NheI site found at 2101 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1048
Illegal EcoRI site found at 1191
Illegal BglII site found at 19 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1048
Illegal EcoRI site found at 1191 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1048
Illegal EcoRI site found at 1191
Illegal NgoMIV site found at 2789
Illegal AgeI site found at 1840 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1073
Illegal BsaI.rc site found at 13
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