Team:Edinburgh/Project

From 2011.igem.org

Revision as of 13:52, 29 June 2011

A project description and safety proposal is supposed to be submitted by July 15.

Project abstract

This year we will create "reactors", 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 two different systems will be investigated:

• Using M13 phage as the scaffold, and attaching enzymes by cell-surface display techniques to the pVIII coat protein.
  • Additionally, it may be possible to attach multiple phage to beads, making a larger "reactor".
• Using bacteria as the scaffold, and attaching enzymes by cell-surface display techniques.
  • This ought to be less efficient, but it may be possible to localise enzymes to the pili or flagella, increasing the synergy...

As an example system, we will try creating E. coli with different cellulase enzymes displayed on the surface, as well as (if possible) M13 phage with the same enzymes displayed on the pVIII coat protein.

Pages

• Phage Reactors
• Cell Display

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>.

• Fuse whatever enzymes to cell-display parts or pVIII or maybe pIII.

•  ???

• Profit!

This was the old Navbox for Edinburgh; now it's obsolete...