September, 01st

Production Team

We wrote the protocol for fecI-fecR* (UPO28*) SDM. It was necessary to remove a PstI site in 1184^thpb (inside of fecR* secuence). We were going to perform two strategies:

• fecI-fecR* SDM to get a composed part.
• fecR* SDM to get only a fecR part. We already had fecI part.

It was not necessary to mutagenize fecA* (UPO8*) or gltD** (UPO17) since we were not going to keep on building devices of them.

Assembly Team

We analyzed all the information we had about parts and devices and took a decision: we were going to do without device 2 (Fec system) and devices 10, 11 and 12 (Glutamate Synthase). The explication was:

• D2 was a control of the sensing system. Moreover we could not synthesize fecA part.
• D10, D11 and D12: glutamate synthase subunits were pretty big so they could give us problems. Also we had problems to remove EcoRI target in one of them by SDM. It was not a huge deal not to continue with that chemoattractant synthesize system because we had other two: aspartate to E. coli and salicilate to P. putida.

We wrote protocols for next devices, SDM of UPO28 and digestion analysis of 1+19 and 16+3.