Composite

Part:BBa_K3369002

Designed by: Qinglin Zeng   Group: iGEM20_Tsinghua   (2020-10-23)


B.subtilis Nitric Oxide synthase with C4-HSL report system


Bacteria nitric oxide synthase (bNOS) from B.subtilis genome was applied as the source of NO. It could utilize L-Asp and oxygen as substrates, receiving the electron from NADPH and producing citrulline and NO. Compared with eukaryote NOS, bNOS lacks reductase domain therefore it requires additional reduction partner to function. Quorum sensing system plays a key role in the formation of bacterial membrane of P.aeruginosa. There are four quorum sensing systems in P.aeruginosa : LasI - LasR and RhlI - RhlR IQS and PQS. Research has shown that the C4-HSL, the QS signal mediated in Rhl-Rhlr system, presented the highest concentration in the extract of the bacterial membrane of P.aeruginosa, indicating C4-HSL its potential as an activation signal switch. Therefore, in our design, we use the Rhl system as our gene switch. When the engineered bacteria enter the bacterial membrane and sense the C4-HSL, it will activate the downstream NOS expression and producing NO.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1070
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1070
    Illegal NheI site found at 18
    Illegal NheI site found at 41
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1070
    Illegal BglII site found at 6
    Illegal BglII site found at 1225
    Illegal BamHI site found at 312
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1070
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1070
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 787
    Illegal SapI.rc site found at 1320


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