Difference between revisions of "Part:BBa K2558212:Design"
 
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===Design Notes===
 
===Design Notes===
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With the hope to find an optimal promotor for our NEON system, we designed 9 mutations on sites 18, 19 and 20 near the luxR binding site and tested one on -10 site that TUST 2017 concluded to show decreased leakage. We conducted experiments to evaluate these lux pR mutants’ reaction to AHL stimulation and their leakage level. The test devices we designed include a constantly expressed luxR and a lux pR (or mutant) (like BBa_K2558211 with original lux pR promotor https://parts.igem.org/Part:BBa_K2558211, and BBa_K2558212 with lux pR-HS promotor https://parts.igem.org/Part:BBa_K2558212).  
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With the hope to find an optimal promotor for our NEON system, we designed 9 mutations on sites -35, -36, -37 near the luxR binding site and tested another on -10 site that TUST 2017 reported to show decreased leakage. We conducted experiments to evaluate these lux pR mutants’ reaction to AHL stimulation and their leakage level. The test devices we designed include a constantly expressed luxR and a lux pR (or mutant) driven sfGFP (like BBa_K2558211 with original lux pR promotor, and BBa_K2558212 with lux pR-HS promotor).
  
  

Latest revision as of 16:14, 17 October 2018


lux pR-HS test device


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1074
    Illegal NheI site found at 1097
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 177


Design Notes

With the hope to find an optimal promotor for our NEON system, we designed 9 mutations on sites -35, -36, -37 near the luxR binding site and tested another on -10 site that TUST 2017 reported to show decreased leakage. We conducted experiments to evaluate these lux pR mutants’ reaction to AHL stimulation and their leakage level. The test devices we designed include a constantly expressed luxR and a lux pR (or mutant) driven sfGFP (like BBa_K2558211 with original lux pR promotor, and BBa_K2558212 with lux pR-HS promotor).


Source

References

Koch B et al. The LuxR receptor: the sites of interaction with quorum-sensing signals and inhibitors. Microbiology. 2005 Nov;151(Pt 11):3589-602.