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Team:Edinburgh/Project
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| - | This year we will create '''"bioreactors"''', consisting of scaffolds for various enzymes, to carry out reactions in an extracellular environment. The hope is that, by combining the activity of multiple synergistic enzymes in a small space, high efficiency will be achieved. At least 4 systems are being considered: | + | This year we will create '''"bioreactors"''', consisting of scaffolds for various enzymes, to carry out reactions in an extracellular environment. The hope is that, by combining the activity of multiple '''synergistic''' enzymes in a small space, high efficiency will be achieved. At least 4 systems are being considered: |
* As a baseline, use bacteria as the scaffold, and attach enzymes by cell-surface display techniques. | * As a baseline, use bacteria as the scaffold, and attach enzymes by cell-surface display techniques. | ||
Revision as of 14:03, 29 June 2011
A project description and safety proposal is supposed to be submitted by July 15.
Project abstract
This year we will create "bioreactors", consisting of scaffolds for various enzymes, to carry out reactions in an extracellular environment. The hope is that, by combining the activity of multiple synergistic enzymes in a small space, high efficiency will be achieved. At least 4 systems are being considered:
- As a baseline, use bacteria as the scaffold, and attach enzymes by cell-surface display techniques.
- As a modified version of the above, use bacteria, but concentrate the enzymes on a small part of it such as the flagella.
- As a fairly novel concept, use M13 phage as the scaffold, and attach enzymes by cell-surface display techniques to the pVIII coat protein.
- As a modified version of the above, attach multiple phage to beads via the pIII protein, making a larger "reactor".
As example systems, we will use cellulases as our enzymes of interest.
Pages
Actions that ought to be carried out
- Make or acquire fusion-ready cellulases (e.g. <partinfo>BBa_K392006</partinfo>, <partinfo>BBa_K392007</partinfo>, <partinfo>BBa_K392008</partinfo>).
- Acquire M13 phage.
- If a simple test system is desired, make fusion-ready amylase? Or something else?
- Make or acquire fusion-ready pVIII gene.
- Perhaps from <partinfo>BBa_K415151</partinfo>.
- Remember it has a leader sequence, coding for: MKKSLVLKASVAVATLVPMLSFA.
- Our final fusion will need the leader sequence followed by the gene of interest followed by pVIII proper.
- Make or acquire fusion-ready pIII gene? (optional)
- See <partinfo>BBa_K415138</partinfo> (complete) or <partinfo>BBa_K257001</partinfo> (sans leader sequence).
- Our final fusion will need the leader sequence followed by the gene of interest followed by pIII proper.
- Make or acquire fusion-ready cell-surface display parts.
- See Berkeley 2009.
- Maybe also see <partinfo>BBa_K265008</partinfo>.
- Make or acquire spacer/linker sequences?
- These are short enough that ordering them as oligos probably makes sense...
- Fuse whatever enzymes to cell-display parts or pVIII or maybe pIII.
- ???
- Profit!
This was the old Navbox for Edinburgh; now it's obsolete...
- Edinburgh 2011
- Project documentation: Project - BioSandwich - Parts - Modeling - Lab Notebook - Safety - Team - Attributions
- Pages for members: Wiki Watch - Useful Links - Sequences - Primers - Practices - Official Profile (has email addresses)
- (edit this navigation box)
