Team:Yale

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<b>Nature’s Antifreeze: Microbial Expression and Characterization of a Novel Insect Antifreeze Protein for De-icing Solutions</b><br /><br />
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            Antifreeze proteins have applications in cryopreservation of food, cells, and organs, as well as in cryosurgery and agriculture. The purpose of this study was to express, purify, characterize, and optimize a novel, hyperactive antifreeze protein recently isolated from the Siberian beetle, Rhagium inquisitor (RiAFP). Large scale (150mg/L), stable production of RiAFP and a RiAFP-GFP fusion protein was achieved in E. coli. Proteins were purified by Ni-NTA affinity chromatography. E. coli expressing RiAFP exhibited increased survival post-freezing. RiAFP inhibited ice recrystallization in a dose-dependent manner. RiAFP also improved tissue morphology of rat livers post-freezing. Preliminary results indicate that RiAFP may have a cryoprotective effect in C. elegans. To optimize the activity of the hypothesized RiAFP binding site, we used directed evolution through multiplex automated genome engineering and are currently screening for mutants with enhanced properties. Finally, to better understand the structure-function relationship, we have generated promising crystals of RiAFP for x-ray crystallography and are now optimizing crystallization conditions.
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welcome to igem yale
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                <p>We demonstrate that the <i>Rhagium inquisitor</i> antifreeze protein possesses great potential for applications requiring freeze resistance or the control of ice growth and morphology.</p>
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    What would it take to make bacteria produce an electrical circuit? One of the most exciting uses for synthetic biology is in the design of biological systems that can replace and improve industrial processes. By achieving industrial goals using biological processes, we predict dramatic reductions in economic and environmental manufacturing costs. Our project is a first step towards biologically synthesized electronic circuits. Based on precedence of naturally redox-capable bacteria, we generated a system in E. coli that reduces metal in solution. Depending on the application, this system has the ability to form a conductive copper sulfide that can be localized with high precision. In the future our bacteria could catalyze metal deposition to form electrical circuits of any desired dimension and complexity.
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                <div id="center"><a href="https://2011.igem.org/Team:Yale/Project"><img src="https://static.igem.org/mediawiki/2011/2/21/YaleBut1.png" /></a></div>
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<a id="nav" href="https://2010.igem.org/Team:Yale/Our Project"> >> to learn more about our project: </a>
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<a id="img-1" href="https://2010.igem.org/Team:Yale/Our Project">Introduction</a>
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about us
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A first year entrant to the iGEM competition, our team is comprised of students from a wide range of backgrounds connected by a common passion for synthetic biology and its applications. After much brainstorming and many wetlab sessions, our team has succeeded in developing a genomic platform for bacterial circuit construction.
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                <p>We expressed, purified, characterized and have begun optimizing the novel, hyperactive <i>Rhagium inquisitor</i> antifreeze protein using a variety of synthetic and molecular biology techniques.</p>
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                <div id="center"><a href="https://2011.igem.org/Team:Yale/Protocols"><img src="https://static.igem.org/mediawiki/2011/a/ac/YaleBut2.png" /></a></div>
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<a id="nav" href="https://2010.igem.org/Team:Yale/Our Team"> >> read more about our team </a>
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                <p>A second year entrant to the iGEM competition, the Yale iGEM team is comprised of students from a wide range of backgrounds connected by a common passion for synthetic biology. </p>
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Latest revision as of 00:53, 29 September 2011

iGEM Yale


Nature’s Antifreeze: Microbial Expression and Characterization of a Novel Insect Antifreeze Protein for De-icing Solutions

Antifreeze proteins have applications in cryopreservation of food, cells, and organs, as well as in cryosurgery and agriculture. The purpose of this study was to express, purify, characterize, and optimize a novel, hyperactive antifreeze protein recently isolated from the Siberian beetle, Rhagium inquisitor (RiAFP). Large scale (150mg/L), stable production of RiAFP and a RiAFP-GFP fusion protein was achieved in E. coli. Proteins were purified by Ni-NTA affinity chromatography. E. coli expressing RiAFP exhibited increased survival post-freezing. RiAFP inhibited ice recrystallization in a dose-dependent manner. RiAFP also improved tissue morphology of rat livers post-freezing. Preliminary results indicate that RiAFP may have a cryoprotective effect in C. elegans. To optimize the activity of the hypothesized RiAFP binding site, we used directed evolution through multiplex automated genome engineering and are currently screening for mutants with enhanced properties. Finally, to better understand the structure-function relationship, we have generated promising crystals of RiAFP for x-ray crystallography and are now optimizing crystallization conditions.

We demonstrate that the Rhagium inquisitor antifreeze protein possesses great potential for applications requiring freeze resistance or the control of ice growth and morphology.

We expressed, purified, characterized and have begun optimizing the novel, hyperactive Rhagium inquisitor antifreeze protein using a variety of synthetic and molecular biology techniques.

A second year entrant to the iGEM competition, the Yale iGEM team is comprised of students from a wide range of backgrounds connected by a common passion for synthetic biology.

Retrieved from "http://2011.igem.org/Team:Yale"