Team 1's Brainstorm for Future Programmed Evolution Research

Barcode of 7 bp (7^4 combinations)

- Use GGA . . . still have to work out restriction enzyme, other details

Changing internal (genetic) elements…riboswitch, other RBS and origins and chaperones

- …Using new ones

- …introducing mutations to improve them

Multistep/more complex pathways?

- Any other metabolic improvements to caffeine -> theophylline pathway

- Stopping the pathway converting ammeline (guanine deamylase), seeing whether that pathway is absolutely necessary to cell survival. (In the melamine -> ammeline -> … pathway)

- What else besides theophylline? (MWSU “laundry list”)

New fitness modules

- Testing /improving /adapting the ThyA fitness module

- Building new fitness modules

New riboswitches

Trying new riboswitch that interacts with coenzyme B12

Building a riboswitch

- …Using/improving Catherine Doyle’s riboswitch builder, which generates potential riboswitch sequences based on a given aptamer sequence

Riboswitch / RBS combo that is a single modular structure (C-Dog?)

Finding a riboswitch that works with ammeline

Our picks

Testing the ThyA fitness module developed at Davidson, or finding other ways to implement it.

Investigating riboswitch-RBS interactions - The problem is that the function of promoters and RBS's are context-dependent, based on each other and on the gene of interest. C-dog is more reliable, but might be improved. The goal of investigating the riboswitch-RBS interactions would be to develop a hybrid riboswitch/C-dog combo, making the riboswitch more reliable.

Testing a new metabolic pathway - choose the pathway based on which riboswitches are already known