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Team:Hawaii/Meeting/2008-06- 5
From 2008.igem.org
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Present: GK, KS, MR, SC, GP, KLS, NW, LG <br> | Present: GK, KS, MR, SC, GP, KLS, NW, LG <br> | ||
| - | + | Presentations:<br> | |
| - | + | *Grace: lux operon night light | |
| - | + | : <b>Discussion Points</b> | |
| - | + | :*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 | *Krystle: bacterial export system, soluble proteins | ||
:: <b>Discussion Points</b> | :: <b>Discussion Points</b> | ||
Revision as of 03:07, 6 June 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Contents |
Agenda
9am St. John 515
- Update progress on experiments performed this week
- competent cell creation (taken 2 days), and testing results
- Part B proposals
- Grace
- Krystle
- 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
- 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.
- 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
- Discussion Points
- 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
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
