Gene Circuit

For more information about the parts used in this circuit click on the parts in the picture that link to the registry.

Click here to see in action how the circuit responds to different inputs.

Our Favorite New Parts

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Used and Characterized Pre-Existing Parts

1. Experience - Promoter (LuxR & 3OC6-HSL regulated), BBa_R0062: the promoter's EC₅₀ could be shown to be dependent on the amount of regulator LuxR (BBa_C0062) in the system by using the three different constitutive promoters BBa_J23100, BBa_J23109, and BBa_J23111 for the production of LuxR. The amount of regulator also demonstrated an influence on the promoter's basal expression or leakiness. Additionally, it could be determined that the promoter can also be activated by the regulators LasR (BBa_C0179) and RhlR (C0171) thus showing crosstalk with these two quorum sensing systems.
   
   
2. Experience - Promoter (LasR & 3OC12-HSL regulated), BBa_R0079: it could be shown that this promoter, which is usually activated by the regulator LasR (BBa_C0179) can also be activated by the regulators LuxR (BBa_C0062) and RhlR (C0171). Therefore, it shows crosstalk with these two quorum sensing systems.
   
   
3. Experience - Promoter (RhlR & C4-HSL regulated), BBa_R0071: the amount of regulator RhlR (C0171) demonstrated an influence on the promoter's basal expression or leakiness as well as on the steepness of the response. This was investigated using two different RBSes controlling the expression level of RhlR, i.e. the RBS B0034 and a RBS optimized for rhlR using the Salis Lab RBS Calculator^[13].
   
   
4. Experience - Regulator LuxR, BBa_R0062: it could be shown, that the regulator LuxR not only activates the promoter BBa_R0062 together with 3CO6-HSL (the product of the enzyme LuxI (BBa_C0161)), but also with 3CO12-HSL (LasI (BBa_C0178) product) and C4-HSL (RhlI (BBa_C0070) product).
   
   
5. Experience - Regulator LasR, BBa_C0179: it could be shown, that the regulator LasR activates the promoter BBa_R0079 mainly together with 3CO12-HSL (the product of the enzyme LasI (BBa_C0178)) and only to a minor extend with 3CO6-HSL (LuxI (BBa_C0161) product) and C4-HSL (RhlI (BBa_C0070) product).
   
   
6. Experience - Regulator RhlR, BBa_C0171: it could be shown, that the regulator RhlR not only activates the promoter BBa_R0071 together with C4-HSL (the product of the enzyme RhlI (BBa_C0070)), but also with 3CO6-HSL (LuxI (BBa_C0161) product) and 3CO12-HSL (LasI (BBa_C0178) product), while the ladder is acting least activating among those three AHL molecules.
   
   
7. Experience - Promoter, BBa_J23100: the constitutive promoter BBa_J23100 was compared to the promoters BBa_J23109 and BBa_J23111 by placing each in a construct in front of the quorum sensing regulator LuxR (BBa_C0062). The system behaved as expected, thus confirming an increasing promoter strength from BBa_J23109 over BBa_J23111 to BBa_J23100.
   
   
8. Experience - Promoter, BBa_J23111: the constitutive promoter BBa_J23111 was compared to the promoters BBa_J23109 and BBa_J23100 by placing each in a construct in front of the quorum sensing regulator LuxR (BBa_C0062). The system behaved as expected, thus confirming an increasing promoter strength from BBa_J23109 over BBa_J23111 to BBa_J23100.
   
   
9. Experience - Promoter, BBa_J23109: the constitutive promoter BBa_J23109 was compared to the promoters BBa_J23100 and BBa_J23111 by placing each in a construct in front of the quorum sensing regulator LuxR (BBa_C0062). The system behaved as expected, thus confirming an increasing promoter strength from BBa_J23109 over BBa_J23111 to BBa_J23100.
   
   
10. Experience - Autoinducer synthetase for the AHL molecule 3OC6-HSL, BBa_C0161: by placing an alginate bead into a millifluidic chip containing sender cells, which are constitutively expressing LuxI (BBa_J2316) in vicinity to a bead containing receiver cells with a construct producing sfGFP in presence of 3OC6-HSL it could be shown that signal transduction was successful. Thus it could be confirmed that the synthetase is producing a quorum sensing molecule.

Characterized New Parts

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