Divide and Conquer Biological Challenges

* We are interested in building the pLac_lux promoter.

If the proteins are always on.....when you have IPTG AND NOT AHL

* We need to get the Lux/AHL system working.

• LuxI
  
• LuxR
  
• lux pR
  
• hybrid plac-lux promoter
  

* We might need to get the Lsr/AI-2 system working.

Our summary of the Lsr system: Davidson/Missouri_Western_iGEM2008#Lsr_.28AI-2.29_cell_signaling_system

• There are NO Lsr parts in the registry:
  
• We will need to make BB parts for:
  

1. LsrR and LsrK which are adjacent to each other in the E. coli genome. We could amplify them together with a total size of over 2600bp.
2. Lsr promoter
3. LuxS that produces DPD that somehow is converted to R-THMF.
4. How do we get cells to make AI-2?

* We need to have an XOR logic gate produced.

Existing Logic Gates

• AND:
  

Existing But Complex AND

Simple But Hypothetical AND
We would need to have:

1. Starts with pTetR
2. over production of LacI^Q repressor but note this link is plain LacI
3. over production of TetR repressor
   
4. Therefore, we will have to build 1) modified promoter and 2) LacI^Q

• NAND:
  

1. I732914

• OR:
  

Simple but Hypothetical OR

• XOR:
  

Simple but Hypothetical XOR

• NOR:
  

1. I732916
   
2. I732917
   

• NOT (Inverters):
  

1. I732205 (NOT/ Dual-repressed NOR)

Self-Contained Inverters

1. J23040 (AHL-dependent inverter)
   

PoPS Output Inverters

1. Q04121 (lac inverter)
   
2. Q04400 (tet inverter)
   

* We need to figure out how to get cells to communicate in a sequence and not stop growing too soon.

• What if Amp^R is secreted and cells are not Amp^R?
  

This would prevent cells down the chain from responding too soon. Erin and Pallavi have conducted experiments with this for both distance and timing.