Team:British Columbia/Results

From 2014.igem.org

(Difference between revisions)
(Created page with "{{:Team:British Columbia/Templates/MainHeader}} <html> <div class="container"> <h1>Results</h1> <p> Sed ut perspiciatis, unde omnis iste natus error sit ...")
 
(18 intermediate revisions not shown)
Line 1: Line 1:
{{:Team:British Columbia/Templates/MainHeader}}
{{:Team:British Columbia/Templates/MainHeader}}
<html>
<html>
 +
 +
<div class="container">
 +
      <h1>Biomining precipitation assays</h1>
 +
 +
Before attempting to demonstrate binding with peptide inserted into the S-Layer, we tested binding with WT cells. After having observed binding, we then hypothesized that the fucose layer in Caulobacter was potentially involved in binding. Since there was a fucose knockout was available, we ere able to confirm fucose binding. Moreover, this observation demonstrated that a precipitation assay was feasible when mixing cells with chalcopyrite.
 +
 +
<html>
 +
<center>
 +
<video width="920" controls>
 +
  <source src="https://static.igem.org/mediawiki/2014/a/a1/UBC_iGEM_chalcopyrite_biomining_1.mp4" type="video/mp4">
 +
Your browser does not support the video tag.
 +
</video>
 +
</center>
 +
</html>
 +
 +
<center>
 +
'''Figure 1.''' Wild type and fucose knockout ''Caulobacter'' cells mixed with Chalcopyrite.
 +
</center>
 +
 +
As a preliminary test, we observed increased Chalcopyrite precipitation with ''Caulobacter'' cells expressing a Chalcopyrite binding peptide in the S-layer. While non-quantitative, the data showed noticeable differences in precipitate settle when the peptide expressed in various locations in the S-layer. See the video below.
 +
 +
<html>
 +
<center>
 +
<video width="920" controls>
 +
  <source src="https://static.igem.org/mediawiki/2014/5/59/UBC_iGEM_chalcopyrite_biomining_2.mp4" type="video/mp4">
 +
Your browser does not support the video tag.
 +
</video>
 +
</center>
 +
</html>
 +
 +
<center>
 +
'''Figure 2.''' Various peptides from Curtis ''et al.'' are displayed in different positions in the S-layer of ''Caulobacter'' cells. The cells were mixed with calcopyrite and precipitate settling was observed. 
 +
</center>
   <div class="container">
   <div class="container">
-
       <h1>Results</h1>
+
       <h1>Biomining protein assays</h1>
 +
 
 +
In an attempt to support the above observation, we incubated ''Caulobacter'' cells with Chalcopyrite and separated the Chalcopyrite by centrifugation. The supernatant and cells adhered to Chalcopyrite (eluted after several buffer washes) were assayed for protein concentration as a read out for biomass. For fucose containing cells and cells expressing Chalcopyrite binding peptides in the S-layer, we expected protein concentration to decrease in the supernatant and increase on Chalcopyrite as compared to controls.
       <p>
       <p>
-
      Sed ut perspiciatis, unde omnis iste natus error sit voluptatem accusantium doloremque laudantium, totam rem aperiam eaque ipsa, quae ab illo inventore veritatis et quasi architecto beatae vitae dicta sunt, explicabo. Nemo enim ipsam voluptatem, quia voluptas sit, aspernatur aut odit aut fugit, sed quia consequuntur magni dolores eos, qui ratione voluptatem sequi nesciunt, neque porro quisquam est, qui dolorem ipsum, quia dolor sit amet consectetur adipisci[ng] velit, sed quia non numquam [do] eius modi tempora inci[di]dunt, ut labore et dolore magnam aliquam quaerat voluptatem. Ut enim ad minima veniam, quis nostrum exercitationem ullam corporis suscipit laboriosam, nisi ut aliquid ex ea commodi consequatur? Quis autem vel eum iure reprehenderit, qui in ea voluptate velit esse, quam nihil molestiae consequatur, vel illum, qui dolorem eum fugiat, quo voluptas nulla pariatur?
+
https://static.igem.org/mediawiki/2014/6/6e/Diagram_biopan_v2_%281%29.png
       </p>
       </p>
   </div>
   </div>
-
</html>
 
 +
<center>
 +
'''Figure 3.''' Bradford assay for protein from cells in supernatant and adhered to Chalcopyrite. Peptides are expressed in wild type ''Caulobacter'' cells.
 +
</center>
 +
<br>
 +
<p>
 +
https://static.igem.org/mediawiki/2014/0/0b/Diagram_bioming2.png
 +
</p>
 +
'''Figure 4.'''Bradford assay for protein from cells in supernatant and adhered to Chalcopyrite.Peptides are expressed in fucose knockout cells.
 +
<p>
 +
https://static.igem.org/mediawiki/2014/2/2b/Diagram4.jpg
 +
</p>
{{:Team:British Columbia/Templates/Footer}}
{{:Team:British Columbia/Templates/Footer}}

Latest revision as of 03:26, 18 October 2014

2014 UBC iGEM

Biomining precipitation assays

Before attempting to demonstrate binding with peptide inserted into the S-Layer, we tested binding with WT cells. After having observed binding, we then hypothesized that the fucose layer in Caulobacter was potentially involved in binding. Since there was a fucose knockout was available, we ere able to confirm fucose binding. Moreover, this observation demonstrated that a precipitation assay was feasible when mixing cells with chalcopyrite.

Figure 1. Wild type and fucose knockout Caulobacter cells mixed with Chalcopyrite.

As a preliminary test, we observed increased Chalcopyrite precipitation with Caulobacter cells expressing a Chalcopyrite binding peptide in the S-layer. While non-quantitative, the data showed noticeable differences in precipitate settle when the peptide expressed in various locations in the S-layer. See the video below.

Figure 2. Various peptides from Curtis et al. are displayed in different positions in the S-layer of Caulobacter cells. The cells were mixed with calcopyrite and precipitate settling was observed.

Biomining protein assays

In an attempt to support the above observation, we incubated Caulobacter cells with Chalcopyrite and separated the Chalcopyrite by centrifugation. The supernatant and cells adhered to Chalcopyrite (eluted after several buffer washes) were assayed for protein concentration as a read out for biomass. For fucose containing cells and cells expressing Chalcopyrite binding peptides in the S-layer, we expected protein concentration to decrease in the supernatant and increase on Chalcopyrite as compared to controls.

Diagram_biopan_v2_%281%29.png

Figure 3. Bradford assay for protein from cells in supernatant and adhered to Chalcopyrite. Peptides are expressed in wild type Caulobacter cells.


Diagram_bioming2.png

Figure 4.Bradford assay for protein from cells in supernatant and adhered to Chalcopyrite.Peptides are expressed in fucose knockout cells.

Diagram4.jpg

© 2014 UBC iGEM

Retrieved from "http://2014.igem.org/Team:British_Columbia/Results"