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Team:Glasgow/Results:fixation
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| - | + | <h1>Fixation Results</h1> | |
| - | + | </br> | |
| + | <h2>Aims</h2> | ||
| + | <p>-To find a suitable material for encapsulation</p> | ||
| + | <p>-To build a construct that will appropriately express this material within a biofilm</p> | ||
| + | <p>-To transform this construct into the chassis and express it</p> | ||
| + | <p>-To characterise the expression and its effects towards fixation and encapsulation of the biofilm</p> | ||
| + | <h2>Methods </h2> | ||
| + | <p>We found our encapsulating material in Capsular Colanic Acid (Chao & Zhang, 2011). For simplicity it was decided to use an existing <a href="http://partsregistry.org/Part:BBa_K200000">colanic acid biobrick</a>, from the Imperial College iGEM 2009 Team.</p> | ||
| + | <p>This biobrick system had been constructed to form a capsule of colanic acid that is anchored to the cell membrane by a ligase> <i>E. coli</i> naturally uses colanic acid to encapsulate itself (Stout & Gottesman, 1990), as shown in the image below. </p> | ||
| + | <div> | ||
| + | <img src="http://partsregistry.org/wiki/images/thumb/a/a3/EM.jpg/650px-EM.jpg"/> | ||
| + | <p><font size="1" color="grey"> Picture 1: EM image of colanic acid encapsulation of <i>E.coli</i>, showing colanic acid anchored to the membrane. (Picture as shown on BBa_K2000000 registry page)</font></p> | ||
| + | </div> | ||
| + | <p>The parts of the colanic acid system that were required to induce overexpression (Gervais, Phoenix & Drapeau, 1992) and export were selected. Anchoring to the membrane was not required. Top10 cells were transformed with the biobricks and the DNA was extracted to begin building a construct. We began attaching Ribosome Binding Sites and terminators to the biobricks we required, using the AlwNI-Method as detailed in our protocols. Due to time constraints we did not finish the entire construct. </p> | ||
| + | <p></p><h4>References</h4> | ||
| + | <p>Chao, Y. & Zhang, T., 2011. Probing Roles of Lipopolysaccharide, Type 1 Fimbria, and Colanic Acid in the Attachment of Escherichia coli Strains on Inert Surfaces.Langmuir. 2011 Sep 20;27(18):11545-53</p> | ||
| + | <p>Gervais FG, Phoenix P, Drapeau GR, 1992. The rcsB gene, a positive regulator of colanic acid biosynthesis in Escherichia coli, is also an activator of ftsZ expression. Journal of Bacterioogy. 1992 Jun ; 174(12): 3964-71</p> | ||
| + | <p>Stout V, Gottesman S, 1990. RcsB and RcsC: a two-component regulator of capsule synthesis in Escherichia coli. Journal of Bacteriology. 1990 Feb ; 172(2): 659-69 </p> | ||
</html> | </html> | ||
Latest revision as of 18:31, 21 September 2011
Fixation Results
Aims
-To find a suitable material for encapsulation
-To build a construct that will appropriately express this material within a biofilm
-To transform this construct into the chassis and express it
-To characterise the expression and its effects towards fixation and encapsulation of the biofilm
Methods
We found our encapsulating material in Capsular Colanic Acid (Chao & Zhang, 2011). For simplicity it was decided to use an existing colanic acid biobrick, from the Imperial College iGEM 2009 Team.
This biobrick system had been constructed to form a capsule of colanic acid that is anchored to the cell membrane by a ligase> E. coli naturally uses colanic acid to encapsulate itself (Stout & Gottesman, 1990), as shown in the image below.
Picture 1: EM image of colanic acid encapsulation of E.coli, showing colanic acid anchored to the membrane. (Picture as shown on BBa_K2000000 registry page)
The parts of the colanic acid system that were required to induce overexpression (Gervais, Phoenix & Drapeau, 1992) and export were selected. Anchoring to the membrane was not required. Top10 cells were transformed with the biobricks and the DNA was extracted to begin building a construct. We began attaching Ribosome Binding Sites and terminators to the biobricks we required, using the AlwNI-Method as detailed in our protocols. Due to time constraints we did not finish the entire construct.
References
Chao, Y. & Zhang, T., 2011. Probing Roles of Lipopolysaccharide, Type 1 Fimbria, and Colanic Acid in the Attachment of Escherichia coli Strains on Inert Surfaces.Langmuir. 2011 Sep 20;27(18):11545-53
Gervais FG, Phoenix P, Drapeau GR, 1992. The rcsB gene, a positive regulator of colanic acid biosynthesis in Escherichia coli, is also an activator of ftsZ expression. Journal of Bacterioogy. 1992 Jun ; 174(12): 3964-71
Stout V, Gottesman S, 1990. RcsB and RcsC: a two-component regulator of capsule synthesis in Escherichia coli. Journal of Bacteriology. 1990 Feb ; 172(2): 659-69
