Team:Hawaii/Meeting/2008-06- 5

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Contents

Agenda

9am St. John 515

  1. Update progress on experiments performed this week
    1. competent cell creation (taken 2 days), and testing results
  2. Part B proposals
    1. Grace
    2. Krystle
    3. Margaret (teleconference in from US mainland)

Minutes

Present: GK, KS, MR, SC, GP, KLS, NW, LG
Presentations:

  • Grace: lux operon night light
Discussion Points
  • rbc may be too strong a promoter for use in the proposed construct
  • test the strength of RBC and explore if it is leaky by creating a lacZ/GFP fusion protein
  • use the fusion protein plasmid from SC, use amplifier to get the RBCL promoter, cut out the fusion site and put into pCC1383. -> 'It can be made into a biobrick if it works.'
  • expression of lux as controlled by lac may not be enough without available lactose
  • question: Is lactose membrane permeable for S. PCC6803? [http://cyano.genome.jp/cgi-bin/cyorf_view.cgi?ORG=syn&ACCESSION=slr1202 YES] PCC6803 encodes two lactose permease proteins, lacF and lacG.
    • may need to find a membrane permeable substance that can bind to the lac promoter
  • Krystle: bacterial export system, soluble proteins
Discussion Points
  • it will be best to focus on pilA and slr2016, signal sequences that have already been experimentally confirmed
  • Margaret: synthetic plasmid biobricks
Discussion Points

Action Items

  • Grace
  • Align PCC6803 rbc promoter with other PCC6803 promoters and other rbc promoters to confirm -10 and -35 consensus sequences
  • What metabolites are needed for luxCDABE expression?
  • What transcription factors/repressors/activators are known to interact with rbc promoter?
  • Can PCC6803 take up IPTG?
  • What is the lifespan of lacI in vivo without degradation signal? With?
  • Look into rbcR
  • Krystle
  • Margaret:
  • Does RSF1010 replicate autonomously in PCC6803?
  • All:
  • Primer and BioBrick orders should be compiled by 6/12

Coming Up


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