IGEM:MIT/2006/Notebook/2007-7-2

=Things to Do=

1. Time Course

9:20- J45120 time zero, 11:20- add salicylic acid, 12:20- extract (3 hrs) (OD600=0.01), 1:20- add SA, 2:20- extract (5 hrs) (OD600=0.21), 3:20- add SA, 4:20- extract (7 hrs) (OD600=1.09), 5:20- add SA, 6:20- extract (9 hrs) (OD600=2.5), 7:20- add SA, 8:20- extract (11 hrs) (OD600=2.5), 9:20- add SA, 10:20- extract (13 hrs) (OD600=2.8)

9:35- J45181 time zero, 11:35- add salicylic acid, 12:35- extract (3 hrs) (OD600=0.01), 1:35- add SA, 2:35- extract (5 hrs) (OD600=0.27), 3:35- add SA, 4:35- extract (7 hrs) (OD600=1.15), 5:35- add SA, 6:35- extract (9 hrs) (OD600=2.0), 7:35- add SA, 8:35- extract (11 hrs) (OD600=3.0), 9:35- add SA, 10:35- extract (13 hrs) (OD600=3.2)

2. Order primers for aroG- NEW PLAN AFTER TALKING WITH JASON- inquired Yale about aroF- aroG+ aroH-, although Pittard suggested it to be doubtful

IDEAL PLAN- they have aroF- aroG+ aroH-, and we can do 2 point mutations on chromosome (Leu8-->Ile8 via T I A at 22 and Ser180--> Phe180 via C I T at 539)

LIKELY PLAN- order aroF- aroG- aroH-, and we can do 3 point mutations on chromosome ((Leu8-->Ile8 via T I A at 22 and Ser180--> Phe180 via C I T at 539 plus a mutation to allow aroG- to gain functionality)

Status- waiting to hear back from Yale

3. Research leucine pathway- Run plasmid plan by Endy/Knight/grad students- DONE

4. Forward emails to other people in labs for GC and Yale- DONE (GC meeting with grad student 2:00 tomorrow, looks good)