IGEM:MIT/2006/Notebook/2007-6-27

Update on Progress

1. Did a 20-hour time course of J45120 and J45181 cultures

2. Have been working with Andreas in the Sauer lab on using HPLC to detect isoamyl acetate

3. Have been collaborating with Professor Tribe in Australia in trying to find a strain which overproduces chorismate (precursor to methyl salicylate)- at present, it seems that we want a aroF and aroG feedback-resistant strain which also has a nonfunctional tyrR gene, but we will discuss this further later

4. Grew J45600 and J45900 cultures up with 15 mM L-leucine added to media: J45900 grew and did not smell, J45600 did not grow

Results from Time Courses

J45181 (Exponential Phase-Sensitive Promoter)

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

1---------------------0.00------------NOT DETECTED

5---------------------0.06------------NOT DETECTED

9---------------------1.86------------16.7

13--------------------2.5-------------16.5

17--------------------3.2-------------2.8

21--------------------2.6-------------0.3

J45120 (Constitutive Promoter)

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

1---------------------0.00------------NOT DETECTED

5---------------------0.00------------NOT DETECTED

9---------------------0.67------------2.3

13--------------------3.3-------------26.9

17--------------------3.3-------------(-0.3)

21--------------------2.8-------------1.1

Last Thing to Do Today

Make overnight 5 mL J45181 culture to construct a growth curve per Francois' suggestion