1. BioWire Progress Report Week Four Orr Ashenberg, Patrick Bradley, Connie Cheng, Kang-Xing Jin, Danny Popper, Sasha Rush

2. Last Week Preliminary tests of LuxI aTc-induced sender reporter Continued building Las, RhI circuits and Lux receiver test constructs Finished CAD design for bacterial stamp Continuous modeling

3. Weiss Circuit (no propagation) tetR promoter LuxI LuxP L LuxRcI LuxP R LuxBox (Lux/CI) hybrid promoter YFP Receiver Sender AHL

4. Propagating Pulse Generator tetR promoter LuxI LuxP L LuxRcI LuxP R Receiver Sender LuxBox (Lux/CI) hybrid promoter YFPLuxI AHL

5. Sender Reporter Test Construct Inducible by aTc (anhydrotetracycline) Expresses eCFP (part #: BBa_E0422) Used GFP filter for preliminary test; CFP expected to arrive last Friday tetR promoter eCFP Sender Reporter Test Construct: aTc in, eCFP out Part BBa_J06003

6. Preliminary pictures of SRTC SRTC + aTc (0.2 µg / mL)SRTC - aTc Oil immersion microscopy, 100x objective, GFP filter

7. Building the Circuits Second round of ligations mostly failed – exposed too long to UV during gel extraction? I0462 (LuxR BioBrick) did not digest properly – Analytical digest worked fine – Repeating digestion as well as picking new colony, in case miniprep of I0462 is bad – May also try to digest differently, in case enzyme is the culprit Repeated on Thursday

8. Photolithography Finished mask design. More?

9. Continuous Model Received MATLAB files from David Braun, which model intracellular pulse generation (but not propagation) Deciphered rate constants (but not units) During downtime, we are extending this model to use positive feedback and to be intercellular rather than intracellular

10. This Week Finish receiver test constructs and begin experiments – J06000 -- receiver should be finished today – J06004 -- receiver w/repressor should be ready in 3 days Finish las test constructs Conduct experiments on SRTC with CFP filter

11. Updated Schedule Week 1 (6/6): Project Choice and Design Week 2 (6/13): Got parts and set up tests Week 3 (6/20): Began building test constructs, finished sender Week 4 (6/27): Finish receiver, receiver w/repressor; CAD a mask Week 5 (7/4): Test component circuits; build stamps, test Week 6 (7/11): Test cell signaling density Week 7 (7/18): Construct IFFL Week 8 (7/25): Test it Week 9 (8/1): “ Week 10 (8/8): “ Week 11 (8/15): “ Week 12 (8/22): “