Device

Part:BBa_K4407014:Design

Designed by: Leyu Xu   Group: iGEM22_Worldshaper-Nanjing   (2022-10-05)


Plac-dps-aceE,Coexpression of dps and aceE in response to lactose


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 368
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 368
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 368
    Illegal BglII site found at 428
    Illegal BglII site found at 2942
    Illegal BglII site found at 3350
    Illegal BamHI site found at 1902
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 368
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 368
    Illegal NgoMIV site found at 3972
    Illegal AgeI site found at 2683
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In our project, we used this device to construct facultative anaerobic Clostridium tyrobutyricum (C. tyrobutyricum). dps protects the bacteria from oxidative damage. It protects DNA against oxidative damage by binding with Fe2+ to prevent it from reacting with H2O2 (Fenton’s reaction). aceE protein is a component of the PDH complex and catalyzes the conversion of pyruvate to acetyl-CoA and CO2. It can function well in aerobic environment. This device combines the function of the two proteins, enhancing viability in aerobic environment by protecting DNA and substituting the main metabolic enzyme PFOR which is inactivated under aerobic condition with PDH in the presence of oxygen. In aerobic environment, we can use lactose to induce dps and aceE expression in the recombinant C. tyrobutyricum to enhance its viability.


Source

Plac is based on BBa_K4119006. dps is based on BBa_K4407001. aceE is based on BBa_K4407002. Terminator is based on BBa_K3585002.

References