Team:Edinburgh/Project

Pages

• Phage Reactors
• Cell Display
• Biorefineries

Designs

The general formats for the basic phage and cell display would be:

• Promoter-RBS-LeaderSequence-Enzyme-(Linker?)-pVIII
• Promoter-RBS-INP-(Linker?)-Enzyme

Actions that ought to be carried out

General

• Make or acquire fusion-ready cellulases (e.g. <partinfo>BBa_K392006</partinfo>, <partinfo>BBa_K392007</partinfo>, <partinfo>BBa_K392008</partinfo>).

• Since a simple test system is desired, make fusion-ready amylase, from E. coli's own malS gene.

• Make or acquire spacer/linker sequences?
  • Short linkers could be ordered as oligos. Berkeley 2009 made some longer linkers which we could order.

Phage display

• Acquire M13 phage.

• PCR from our [http://www.neb.com/nebecomm/products/productn4018.asp M13mp18] to get:
  • pVIII mature protein
  • pVIII leader sequence, coding for: MKKSLVLKASVAVATLVPMLSFA.

• 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.

Cell display

• Make or acquire fusion-ready cell-surface display parts.
  • See Berkeley 2009 (but perhaps ignore them).
  • Instead, <partinfo>BBa_K265008</partinfo> (ice nucleation protein) looks promising.
  • The fliC gene seems to be the target for flagella display; note that the displayed protein has to be inserted into the middle of it.

Biorefinery

To add...

Completion

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

•  ???

• Profit!