Cyanobacteria Oscillator E. Coli Harvard iGEM 2006 Week 3 Peng, David, Jeff, Hetmann

Overall goal We aim to create a kaiABC + reporter BioBrick which expresses a circadian rhythm in both E. coli and cyanobacteria. (We are still brainstorming ideas of what to do; this week was review literature / grow strains / contact people)

Blue = biobricks cut site Green = can be obtained from cyanobacteria Prof. Susan Golden and others have this

We’ve obtained three strains of cyanobacteria: From Eric Webb at Woods Hole: WH8102, a marine strain We’re probably not going to use it because it’s reputed to be very difficult to grow and modify From Peter Weigele at MIT: PCC7942 and PCC6803, both freshwater strains PCC6803 can metabolize glucose PCC7942 is the most common strain we’ve encountered in the literature on cyanobacteria circadian oscillation Both strains’ genomes have been sequenced Both can be reliably transformed by exogenous DNA

Incubator We modified an incubator by installing a timer-controlled fluorescent light (32W + 22W bulbs) The lights are set to stay on for 16 hours each day According to Peter Weigele, we should expect growth in 5-7 days (Tuesday-Thursday this week)

Next steps Design primers for extracting KaiABC Learn protocols for transforming PCC7942 and PCC6803 (Maybe): synthesis of kaiABC Waiting to hear back from… Susan Golden at UT Austin Alexander van Oodenaarden at MIT Jeffrey Chabot