Team:HokkaidoU Japan/Kill switch

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Team:HokkaidoU Japan

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• Top
• Project
  • Overview
    • Project Description
    • Background
    • Proof of concept
    • Modeling
    • Achievements
  • MULTIMERIZATION
    • Overview
    • Methods
    • Results
    • Conclusion
  • FUTURE WORK
    • Circularization
    • Kill Switch
    • Transport
    • Aggregation
  • Extra Materials
    • Parts
    • Notebook
    • Protocols
    • Safety
• Human practices
• Collaborations
• Team
  • Attribution
  • Member

Fig. 1. Expected effect of the new cell-death system as an application of SAP

We came up with an idea of utilizing SAP for cell death system. To be specific, we expect SAP which is excessively accumulated inside of E.colii would be vital damage for E.colii by disturbing its metabolism or catabolism, for example. In addition, this system is expected to cause cell death without cell lysis, which is a desirable feature when you want to handle E.colii containing harmful substances like heavy metal ion inside.

For our new cell death system, we designed a construct shown in fig.2. The construct has a lactose promoter upstream of RBS and SAP coding region, thus we can control the timing of switching-on the device. We ordered the parts of DNA from IDT, and after subcloning, we put the piece of DNA on pSB1C3 vector and transformed it into E.coli.

Fig. 2. The construct for kill-switch system

Unfortunately we didn?ft have enough time to assess this cell death system, but we planned to assess the effect of this biodevice according to the following protocol;

1. IPTG induction
2. Incubation at 37?°C
3. Measurement of OD600 every hour
4. If the system works correctly, OD600 after IPTG induction should decrease every hour, whereas the negative controls wouldn?ft show apparent decrease in OD600.

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