Team:Edinburgh/Project

From 2011.igem.org

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==Project abstract==
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__NOTOC__
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'''This page is obsolete. The project should be documented at the relevant subpages:'''
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: ''To be written''
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* [[Team:Edinburgh/Phage Reactors 2.0|Phage Reactors]]
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* [[Team:Edinburgh/Cell Display|Cell Display]]
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* [[Team:Edinburgh/Biorefineries|Biorefineries]]
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==Project details==
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==Designs==
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: ''To be written''
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The general formats for the basic phage and cell display would be:
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{{:Team:Edinburgh/Template:Navbox}}
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* Promoter-RBS-LeaderSequence-Enzyme-(Linker?)-pVIII
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* Promoter-RBS-INP-(Linker?)-Enzyme
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==Actions that ought to be carried out==
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===General===
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* Make or acquire fusion-ready cellulases (e.g. <partinfo>BBa_K392006</partinfo>, <partinfo>BBa_K392007</partinfo>, <partinfo>BBa_K392008</partinfo>).
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* Since a simple test system is desired, make fusion-ready amylase, from ''E. coli's'' own ''malS'' gene.
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* Make or acquire spacer/linker sequences?
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** Short linkers could be ordered as oligos. [[Team:Edinburgh/Berkeley 2009 Parts|Berkeley 2009 made some longer linkers]] which we could order.
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===Phage display===
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* Acquire M13 phage.
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* PCR from our [http://www.neb.com/nebecomm/products/productn4018.asp M13mp18] to get:
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** pVIII mature protein
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** pVIII leader sequence, coding for: MKKSLVLKASVAVATLVPMLSFA.
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* Make or acquire fusion-ready pIII gene? (optional)
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** See <partinfo>BBa_K415138</partinfo> (complete) or <partinfo>BBa_K257001</partinfo> (''sans'' leader sequence).
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** Our final fusion will need the leader sequence followed by the gene of interest followed by pIII proper.
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===Cell display===
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* Make or acquire fusion-ready cell-surface display parts.
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** See [https://2009.igem.org/Team:Berkeley_Wetlab/Cell_Surface_Display_Parts Berkeley 2009] (but perhaps ignore them).
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** Instead, <partinfo>BBa_K265008</partinfo> (ice nucleation protein) looks promising.
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** 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.
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===Biorefinery===
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:''To add...''
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===Completion===
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* Fuse whatever enzymes to cell-display parts or pVIII or maybe pIII.
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* ???
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* '''Profit!'''
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Latest revision as of 14:41, 28 July 2011

This page is obsolete. The project should be documented at the relevant subpages:

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.

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!