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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You are provided with this team page template with which to start the iGEM season.  You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki. You can find some examples <a href="https://2009.igem.org/Help:Template/Examples">HERE</a>.
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You <strong>MUST</strong> have a team description page, a project abstract, a complete project description, a lab notebook, and a safety page.  PLEASE keep all of your pages within your teams namespace. 
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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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                <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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                <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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                <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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                <div id="center"><a href="https://2011.igem.org/Team:Yale/Team"><img src="https://static.igem.org/mediawiki/2011/0/02/YaleBut3.png" /></a></div>
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{|align="justify"
 
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|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
 
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|[[Image:Yale_logo.png|200px|right|frame]]
 
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''Tell us more about your project.  Give us background.  Use this as the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
 
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|align="center"|[[Team:Yale | Team Example]]
 
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{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
 
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!align="center"|[[Team:Yale|Home]]
 
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!align="center"|[[Team:Yale/Team|Team]]
 
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!align="center"|[https://igem.org/Team.cgi?year=2011&team_name=Yale Official Team Profile]
 
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!align="center"|[[Team:Yale/Project|Project]]
 
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!align="center"|[[Team:Yale/Parts|Parts Submitted to the Registry]]
 
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!align="center"|[[Team:Yale/Modeling|Modeling]]
 
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!align="center"|[[Team:Yale/Notebook|Notebook]]
 
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!align="center"|[[Team:Yale/Safety|Safety]]
 
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!align="center"|[[Team:Yale/Attributions|Attributions]]
 
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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"