IGEM:MIT/2006/Notebook/2007-7-3

Things to Do

1. Request Japanese plasmid which overexpresses leucine in E. coli- DONE

2. Ask Professor Prather if she thinks that leucine is limiting in our reaction- DONE

3. Perform experiment that Reshma suggested (add .02 g/L of leucine to J45600 and J45900 cultures)

4. Resend request to Yale for aroF- aroG- aroH-, make sure it gets ordered today- DONE (AB3257 (aroF- aroG- aroH-) is on its way in today's mail!!!)

5. Ask Endy if they think that it is a good idea to add new promoters/RBSs to precursor devices in parallel with obtaining overexpressing strains/what overall goals should be a priority- DONE

6. Run GC samples from yesterday's time course- WORKING ON IT

7. Meet with Alex about GC (2 PM)- DONE (need to prepare isoamyl acetate in heptane and heptane extraction of LB media, Alex seemed optimistic)

8. Print out GC procedure for isoamyl acetate for meeting- DONE

Results from Time Courses

J45120 (Constitutive Promoter)

Time (Hrs)------------OD600-----------Methyl Salicylate (ppm)

3---------------------0.01------------NOT DETECTED

5---------------------0.21------------(-0.4)

7---------------------1.09------------2.6

9--------------------2.5--------------16.9

11--------------------2.5-------------

13--------------------2.8-------------

J45181 (Exponential-Phase Sensitive Promoter)

Time (Hrs)------------OD600-----------Methyl Salicylate (ppm)

3---------------------0.01------------NOT DETECTED

5---------------------0.27------------(0.1)

7---------------------1.15------------6.5

9--------------------2.0--------------

11--------------------3.0-------------

13--------------------3.2-------------