Part:BBa_K2116011
AND gate regulated by Nitric Oxide and 3OC6HSL
Promoter norV is the native promoter controlling the nitric oxide reduction operon (norRVW) in E. Coli. [1] It's transcriptional regulator, NorR, can sense nitric oxide and activate gene expression.
EsaR is a transcriptional regulator of the P. stewartii quorum sensing system. Unlike other quorum sensing regulators, it acts as a repressor and not an activator. It binds DNA through an 18bp binding site called esabox. When bound to 3OC6HSL, it is released to allow transcription.
We created an AND gate by placing one esabox as roadblock, and one esabox right before the first norR binding site of the PnorV promoter. This second esabox was added to create competitive binding between norR and esaR.
We have characterised the AND gate using constitutively expressed esaR on a medium copy plasmid (pRB322/rop). We rely on the native NorR production in E.coli to activate the AND gate.
[[File:Bba K2116011 AND gatebehaviour.png|500px|thumb|left|The AND gate was tested in presence of DETA/NO, which releases NO, and 3OC6HSL. Activation was quantified through GFP expression. The AND gate works as expected, reaching maximal expression only when both NO and AHL are present at the highest concentration tested.]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
None |