Team:UIUC-Illinois/Project/Design

From 2012.igem.org

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       <td>This is a theoretically negative fluorescence control due to the endonuclease activity of a Control Binding Site bound PUF-PIN protein silencing the YFP gene.</td>
       <td>This is a theoretically negative fluorescence control due to the endonuclease activity of a Control Binding Site bound PUF-PIN protein silencing the YFP gene.</td>
     </tr>
     </tr>
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<tr>mCherry + Protet
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<td>In order to minimize confounding factors coming from the YFP reporter itself, we tested our construct with a replaced RFP reporter as well</td></tr>
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<tr>mCherry + Protet + PUF-PIN
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<td>This test was to test the effects of the PUF-PIN endonuclease activity on a reporter other than YFP to pinpoint possible problems stemming from a YFP reporter</td>
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</tr>
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   </tbody>
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Revision as of 19:01, 29 September 2012

Header

Project Design

Project Design

  • Overview
  • PUF+PIN Fusion
  • YFP Reporter
  • Non-Specific Control
  • Reliability experiments
  • Theoretical Results
  • Experimental Design


    In designing our project we based our quantitative tests on fluorescence measured by a plate fluorescence reader. Our constructs were created in ways that best suit providing evidence for our hypothesis of the PUF-PIN fusion protein showing endonuclease activity. Generally, our results were collected from quantifying two main PUF-PIN fusion protein types, a wild type and a mutant type, in different conditions. These two fusion proteins had recognition sites that differed by one base pair.

    Click on the list to the left to read about each of our constructs and why we decided to do them. All expressions were done in vivo with the DH5a strain of E.Coli on pBAD30 or Protet plasmids.

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