1 / 11

BioWire Progress Report Week Four

BioWire Progress Report Week Four. Orr Ashenberg, Patrick Bradley, Connie Cheng, Kang-Xing Jin, Danny Popper, Sasha Rush. Preliminary tests of LuxI aTc-induced sender reporter Continued building Las, RhI circuits and Lux receiver test constructs Finished CAD design for bacterial stamp

nakia
Download Presentation

BioWire Progress Report Week Four

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

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

  2. 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 Last Week

  3. tetR promoter LuxI LuxPL LuxPR LuxR cI LuxBox (Lux/CI) hybrid promoter YFP Weiss Circuit (no propagation) Sender AHL Receiver

  4. LuxPL LuxPR LuxR cI LuxBox (Lux/CI) hybrid promoter YFP LuxI Propagating Pulse Generator tetR promoter Sender LuxI AHL Receiver

  5. tetR promoter eCFP Sender Reporter Test Construct: aTc in, eCFP out Part BBa_J06003 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

  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): “

More Related