Difference between revisions of "Team:UGent Belgium/Biofunction"

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<h2> Biofunction </h2>
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<h4> Overview </h4>
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<p class="justify">With the biofunction group we will make several constructs to test and validate our system. We will try to enhance the function of the 3D printed shape by using biological nucleation proteins. In this way, we can improve the condensation capacity of the water collector. To achieve this, we will use the InaZ gene, an ice nucleating protein (INP) of <i>Pseudomonas syringae</i>. These INP’s are known to cause ice damage on plants and are also frequently used in snowmakers. Recently however, <i>Pseudomonas syringae</i> was also found in clouds, where they can maybe help in rain formation [1-3].</p>
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<p class="justify">The schematic structure of INP can be found underneath. It consists of a membrane binding N-terminal domain, some internal repeating domains responsible for the nucleation, and a C-terminal domain.</p>
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<div class="center"><img src="https://static.igem.org/mediawiki/2016/a/a0/T--UGent_Belgium--INP_structure.jpg" alt="INP structure"></div>
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<p>On the water collector, biotin will be coated. Hence, our constructs must contain streptavidin for effective binding this to the 3D shape. To produce and attach these INP’s to our water collector, we will investigate 2 options:
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  <li>Production of an INP-streptavidin fusion protein in <i>E. coli</i>, followed by lysis of the cells and extraction of the fusion-protein</li>
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  <li>Separate membrane expression of both streptavidin and INP in <i>E. coli</i></li>
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Revision as of 20:58, 29 August 2016

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Biofunction

Overview

With the biofunction group we will make several constructs to test and validate our system. We will try to enhance the function of the 3D printed shape by using biological nucleation proteins. In this way, we can improve the condensation capacity of the water collector. To achieve this, we will use the InaZ gene, an ice nucleating protein (INP) of Pseudomonas syringae. These INP’s are known to cause ice damage on plants and are also frequently used in snowmakers. Recently however, Pseudomonas syringae was also found in clouds, where they can maybe help in rain formation [1-3].

The schematic structure of INP can be found underneath. It consists of a membrane binding N-terminal domain, some internal repeating domains responsible for the nucleation, and a C-terminal domain.

INP structure

On the water collector, biotin will be coated. Hence, our constructs must contain streptavidin for effective binding this to the 3D shape. To produce and attach these INP’s to our water collector, we will investigate 2 options:

  1. Production of an INP-streptavidin fusion protein in E. coli, followed by lysis of the cells and extraction of the fusion-protein
  2. Separate membrane expression of both streptavidin and INP in E. coli