Team:Edinburgh/Project

From 2011.igem.org

(Difference between revisions)

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This page is obsolete.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!

@@ Line 1: / Line 1: @@
-==Project abstract==
+{{: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+'''This page is obsolete.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:'''
-: ''To be written''
+* [[Team:Edinburgh/Phage Reactors 2.0|Phage Reactors]]
+* [[Team:Edinburgh/Cell Display|Cell Display]]
+* [[Team:Edinburgh/Biorefineries|Biorefineries]]
-==Project details==
+==Designs==
-: ''To be written''
+The general formats for the basic phage and cell display would be:
-{{:Team:Edinburgh/Template:Navbox}}
+* 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. [[Team:Edinburgh/Berkeley 2009 Parts|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 [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.
+* ???
+* '''Profit!'''
+</div> <!-- /main_body-->
+<html></div> <!-- /mids --></html>