Team:Edinburgh/Project
From 2011.igem.org
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- | + | {{:Team:Edinburgh/tech/Navbox}} | |
+ | <html><script type="text/javascript" >$(document).ready(function() { | ||
+ | getMenus('home', 'home_project'); | ||
+ | }); </script></html> | ||
+ | <div class="main_body"> | ||
+ | __NOTOC__ | ||
+ | '''This page is obsolete. The project should be documented at the relevant subpages:''' | ||
- | + | * [[Team:Edinburgh/Phage Reactors 2.0|Phage Reactors]] | |
+ | * [[Team:Edinburgh/Cell Display|Cell Display]] | ||
+ | * [[Team:Edinburgh/Biorefineries|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== | ==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>). | * 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. [[Team:Edinburgh/Berkeley 2009 Parts|Berkeley 2009 made some longer linkers]] which we could order. | ||
- | + | ===Phage display=== | |
- | + | ||
- | + | ||
- | + | ||
- | * Make or acquire fusion-ready pIII gene? | + | * 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). | ** 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. | ** 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. | * Make or acquire fusion-ready cell-surface display parts. | ||
- | ** See [https://2009.igem.org/Team:Berkeley_Wetlab/Cell_Surface_Display_Parts Berkeley 2009]. | + | ** See [https://2009.igem.org/Team:Berkeley_Wetlab/Cell_Surface_Display_Parts 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. | * Fuse whatever enzymes to cell-display parts or pVIII or maybe pIII. | ||
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* ??? | * ??? | ||
- | * Profit! | + | * '''Profit!''' |
- | + | </div> <!-- /main_body--> | |
+ | <html></div> <!-- /mids --></html> |
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.
- 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!