Ramping up Programmed Evolution

1) Riboswitch

• Dual ligand
• Turning on and off riboswitch

2) Increased variations in the module including the chaperones

• Increase in variation within current categories. (Copy number, promoter strength)
• New category: mutation of individual base pairs
• Controlling interaction of riboswitch and aptamer

3) Ecoli-yeast

• Temperature difference
• Simple vs. complex pathway of translation

4) The number of steps in the metabolic pathway

• The formation of byproducts
• More than one compound of interest, possibly by decomposition.
• The number of steps to reach differences in significant results.

5) Introducing mutations like in the promoter region, the ribosome binding site, elements found in nature like nickel, cadmium 6) Utilize other bacterial species i.e. B.subtilius