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Team:Hawaii/Meeting/2008-06- 5
From 2008.igem.org
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== Minutes == | == Minutes == | ||
Present: GK, KS, MR, SC, GP, KLS, NW, LG <br> | Present: GK, KS, MR, SC, GP, KLS, NW, LG <br> | ||
| - | Presentations:<br> | + | *Presentations:<br> |
| - | Grace: lux operon night light | + | :*Grace: lux operon night light |
| - | : <b>Discussion Points</b> | + | :: <b>Discussion Points</b> |
| - | :*rbc may be too strong a promoter for use in the proposed construct | + | ::*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 | + | ::*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.' | + | :::*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 | + | ::*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. | + | :::**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 | + | :::**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> |
| - | :*it will be best to focus on pilA and slr2016, signal sequences that have already been experimentally confirmed | + | ::*it will be best to focus on pilA and slr2016, signal sequences that have already been experimentally confirmed |
| - | Margaret: synthetic plasmid biobricks | + | *Margaret: synthetic plasmid biobricks |
| - | : <b>Discussion Points</b> | + | :: <b>Discussion Points</b> |
== Action Items == | == Action Items == | ||
Revision as of 03:05, 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
- 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 
]
