Team:Arizona State/Parts
Parts
Overview of Parts
Our team submitted a total of 6 parts to the iGEM registry. The parts that were submitted were all components to N-acyl homoserine lactone (AHL) quorum sensing systems. The parts are all RFC10, 23, and 1000 compatible with some parts possessing other compatibilities. There are 5 basic parts that are part of the part collection and 1 composite part. The senders for the Aub, Bja, Bra, Cer, and Sin systems were submitted as part of the inductance test with the F2620 part, and compose a submitted part collection. The 1 composite part is the modular cassette (vector) for the inducer.
Our Favorite Parts
BBa_K2033000:N-dodecanoyl-L-homoserine lactone (C(12)-HSL) Sender- AubI
BBa_K2033011:N-Acyl Homoserine Lactone (AHL) Modular Sender Vector
Quorum Sensing-F2620 Inductions
The part Bba_F2620 (found here), designed by Barry Canton and Anna Labno from MIT, is a device designed to output PoPs when LuxR is activated. This was used by the ASU team to test interactions between inducers from other quorum sensing systems. The Aub, Bja, Bra, Cer, and Sin systems stem from different organisms and their inducers were submitted as parts to the registry with the purpose of completing this induction test. With very few Senders currently found in the registry (only around 7), the addition of 5 Senders adds significant depth to the Sender pool.
Each Sender may also be referred to as an AHL synthase, as it produces a specific AHL. The 3D structures of the AHLs produced by the systems we examined are found in the Project Description. The induction test was done on a 96-well plate, which was run in a plate reader to read GFP expression levels. Aside from the controls, two different AHL concentrations were used. The AHL source that was used was filtered out via liquid-liquid extraction and then re-seeded with new cells. This allowed an 8-hour read to produce a definitive trend in the growth curve, providing information about the relationship of these Senders with F2620.
