Team:Imperial College London/M2/EncapsulationRationale

From 2009.igem.org

(Difference between revisions)
(The need for encapsulation:)
(Phase 1:)
Line 7: Line 7:
{{Imperial/09/Division}}
{{Imperial/09/Division}}
-
==<b>Phase 1:</b>==
 
-
 
-
Our rationale for looking for natural sources of acid resistance is that it is easier to hack existing pathways than to transfer large numbers of genes into a different chassis with a dissimilar genetic background.
 
-
Based on natural sources of acid resistance, <i>Lactobacillus</i>, <i>E.coli</i> and <i>B.subtilis</i> were shortlisted as potential chassis.
 
-
 
-
Of these three organisms, <i>E.coli</i> was chosen as it is safe, easy to work with and possesses a broad range of acid resistance strategies.
 
-
 
-
 
-
 
-
 
-
{{Imperial/09/Division}}
 
==<b>Phase 2:</b>==
==<b>Phase 2:</b>==

Revision as of 21:44, 19 October 2009

II09 Thumb m2.pngModule 2 - Acid Resistance Overview

II09 TimelineM2.png



Phase 2:

We hacked E.coli’s acid resistance module in three places to achieve the production of a safe acid resistant capsule.

Acid Resistant Polymer – Colanic acid: E.coli naturally produces a harmless acid–resistant polymer known as colanic acid. Colanic acid is a polymer of glucose, galactose and glucuronic acid. By tapping into the pathway that initiates colanic acid biosynthesis, we can turn on its production via the modulation of a transcription factor encoded by a gene called RcsB.

 About RcsB


Safety – Biofilm prevention: In nature, colanic acid acts as a binding agent between individual cells over which a biofilm can be formed. While colanic acid itself is harmless, biofilm formation is associated with a number of virulence factors. To prevent biofilm formation from occurring, we have tapped into a second pathway such that our cells become locked into colanic acid production. The gene responsible for preventing biofilm formation is a transcription factor encoded by a gene called YgiV.

  About YgiV


Microencapsulation – Colanic acid tethering: In nature, colanic acid is associated with but not attached to the cell surface. To facilitate whole cell encapsulation, we have modified a third pathway to fix the colanic acid to the surface of the cell. This involves the over–production of an enzyme called Rfal.

  About Rfal



Mr. Gene   Geneart   Clontech   Giant Microbes