IAP 2004:Aspartate Sender

From 2006.igem.org

Aspartate Sender

Devices

Attract.1 (I3200) The AspA attractant sender device takes TIPS in from the hk022cI switch output ON device (I3410) and creates AspA. AspA aminates fumarate to make aspartate. Aspartate can be used as a bacterial chemotaxis signal. Once a cell has had its switch turned ON, it synthesizes and excretes aspartate to attract neighboring cells. For this mechanism to function, our system is placed in E. coli cells that are AspA- mutants.

This device is part of a polycistronic message, so the TIPS flow straight through to the CFP reporter device (E0422).

aspA CDS (C0083) The start codon was changed from GTG to ATG to match BioBricks standards. Five silent mutations were introduced to eliminate EcoRI and PstI restriction enzyme sites. EcoRI sites were eliminated at bases 588 and 657 with C -> T mutations. PstI sites were eliminated at bases 624, 999, and 1125 with G -> A mutations. The mutations were designed to match codons with similar frequencies of usage in E. coli.

Report.1 (E0422) The CFP reporter device takes TIPS in from the hk022cI switch output ON device (I3410) and creates CFP. Glowing cells in our system will be those that are in the ON state and are synthesizing AspA.