Team:UCSF/ProjectOverview

From 2011.igem.org

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<regulartext>For our project, we decided to utilize the a-agglutinin half of the aggregation display system to form artificial biofilms. By placing sequences of strong adhesive proteins behind the GPI anchor-Aga1-Aga2 gene and overexpressing them, we were able to create yeast cells that could adhere to different surfaces and even other yeast cells. A couple genes we used to attach to the end of the Aga1p-Aga2p complex were mice cadherins, proteins that mussels used to stick to rocks, and proteins used by other yeast species to initiate biofilm formation.  <p></regulartext>
<regulartext>For our project, we decided to utilize the a-agglutinin half of the aggregation display system to form artificial biofilms. By placing sequences of strong adhesive proteins behind the GPI anchor-Aga1-Aga2 gene and overexpressing them, we were able to create yeast cells that could adhere to different surfaces and even other yeast cells. A couple genes we used to attach to the end of the Aga1p-Aga2p complex were mice cadherins, proteins that mussels used to stick to rocks, and proteins used by other yeast species to initiate biofilm formation.  <p></regulartext>
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<img align="left" style="margin-bottom:0px; width: 350px; margin-top:0px; padding:0;" src="http://dl.dropbox.com/u/24404809/Surface%20Display%20Figure1.jpg">
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<regulartext1> <b><br><BR><BR><BR><BR><br><BR><BR><BR><BR><BR><bR><BR><BR><BR><BR>Figure 1:</b> Control, non-induced mgfp-
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5 yeast. Dispersed, no major
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clustering. </regulartext1>
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<img align="left" style="margin-bottom:0px; width: 350px; margin-top:0px; padding:0;" src="http://dl.dropbox.com/u/24404809/Heterodimers.jpg">
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<BR>
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<regulartext1> <b><br><BR><BR><BR><BR><br><BR><BR><BR><BR><bR><BR><BR><BR><BR>Figure 2:</b> Experiment, induced mgfp-
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5 yeast. 3-D cluster of yeast cells
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growing around and on top of other
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cells. </regulartext1>
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<div id="right1">
<div id="right1">
<img style="margin-bottom:0px; width:500px; margin-top:0px; padding:0;" src="http://dl.dropbox.com/u/24404809/Surface%20Display%20Figure1.jpg">
<img style="margin-bottom:0px; width:500px; margin-top:0px; padding:0;" src="http://dl.dropbox.com/u/24404809/Surface%20Display%20Figure1.jpg">
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<regulartext1> <b>Yeast Cell Surface Display</b>  </regulartext1>
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<regulartext1> <b>Yeast Cell Surface Display</b>  </regulartext1> </div>
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<regulartext1> <b>Yeast Cell Surface Display</b>  </regulartext1>
<regulartext1> <b>Yeast Cell Surface Display</b>  </regulartext1>

Revision as of 22:10, 27 September 2011

introduction
the team
our project
parts
requisites
attributions
extras

Queen's

Background

Agglutination, a type of aggregation specific to mating in Sacchromyces cerevisiae, occurs with two surface proteins: alpha-agglutinin and a-agglutinin. Alpha-agglutinin is produced only by alpha-cells and a-agglutinin is produced only by a-cells. During mating, each cell releases pheromones and accepts pheromones secreted by other cells. The alpha-cells respond to a-cell pheromones while the a-cells respond to alpha pheromones. When each cell has received enough of the correct molecule, it will begin to make its agglutinin. In close proximity, the alpha-agglutinin and the a-agglutinin will aggregate, causing cell-cell adhesion. The two yeast cells will come close enough and eventually fuse into one diploid cell.

We took a closer look into the protein, a-agglutinin, and found that it was composed of two subunits: Aga1p and Aga2p. The Aga1 and Aga2 genes lie under a promoter that switches on when the cell has received pheromones from a mate. When the proteins are expressed, Aga1p is attached to the cell surface by a GPI anchor and is also connected to Aga2p through disulfide bonds.

For our project, we decided to utilize the a-agglutinin half of the aggregation display system to form artificial biofilms. By placing sequences of strong adhesive proteins behind the GPI anchor-Aga1-Aga2 gene and overexpressing them, we were able to create yeast cells that could adhere to different surfaces and even other yeast cells. A couple genes we used to attach to the end of the Aga1p-Aga2p complex were mice cadherins, proteins that mussels used to stick to rocks, and proteins used by other yeast species to initiate biofilm formation.

















Figure 1:
Control, non-induced mgfp- 5 yeast. Dispersed, no major clustering.
















Figure 2:
Experiment, induced mgfp- 5 yeast. 3-D cluster of yeast cells growing around and on top of other cells.
Yeast Cell Surface Display
Yeast Cell Surface Display