IGEM:MIT/2005/Wednesday 7/20 Experiment Meeting "Minutes"

Experimental Planning Meeting:

PURPOSE: Do run through of experimental meeting What do you have What do you need

INPUT

Maxine:

1. Permeablizing membrane -- like making cells competent
2. Attach FFK peptide to antisense agents (RNA) -> lets antisense agent enter cell -> proteins to nucleic acid -> Drew knows things -- proteins that splice out ? -- catch splicing process -- make a tadpole -- protein head and oligo tail
3. mutant cells: actinomycin (anti biotic) -- strain that takes big things, or just actinomycein?
4. DNA transport: what exists: conjugation? comEC system (B. subtlis)

Will:

1. Tests this morning didn't go so well
2. Grew Mc4100 in M9 (doesn't fluoresce), LB
   • no pellet in M9 solution
   • LB cells resuspended in flur. diluted in M9 media
3. Severe contamination -- Will != sterile
4. Do the whole set of tests again (can do MC4100 when I do overnights)
5. Kate says: this expt. was planned at the end of the day, and that was no good at all.
6. Would like to start dealing with OligoFlurs. --> need assembly

HEAD UNIT

Jenny

1. scFv's moving forward

TOXR

Jenny:

1. MalE and PhoA -- PhoA had multiple PCR products: TK says raise annealing temperature; Kate: Blast primers against e. coli genome. Though we can just excise band and use that, it makes sense to go forward and do it again

TK:

1. Took protein sequences from cholera toxR -- vibrio species have similar genes, v. fischeri -- also serratia marcescens (don't really believe it is there) -- (sprayed over San Fran)
2. ATCC -- cholera dna is BL1
   • go ahead and get it -- use this as an exercise
3. Annotation of translation start on cholera toxR was initially wrong -- there is ambiguity -- ack!! We need to check what we ordered!!

FECA

Annie

1. track down fur- strain -- hoping to receive -- how do we get it? RAY = Point of contact
2. jen and annie talking with samantha to work out recombineering
3. where do we fuse our scFv? 3 domains: 7 fusions for one domain alone -- working out details -- having a team meeting about this at 4.30 -- should be a team decision -- can we do it in parallel?
4. FecA in process, FecA prom -- know by end of day tom. if need new primers
5. FecR and FecI starting pcr

SIGNAL PROCESSING:

Ray:

1. Testing parts, RNA based regulation from IAP 2003

ACTUATOR:

Jen:

1. waiting on promoters, going to assemble with gfp when those are ready

Side note: lets talk about risk and danger! Side note: are we overextending ourselves? -gathering materials != doing expt -- might as well gather materials

TO DO: Look into B. Subtleis