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

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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