BerkiGEM2007Present2

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  <p><a href="https://2007.igem.org/Berkeley_UC">&lt;&lt;&lt; Return to UC Berkeley iGEM 2007 </a></p>
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  <p><a href="https://2007.igem.org/BerkiGEM2007Present5">&lt;&lt;Previous Section: Growth Control</a> | <a href="https://2007.igem.org/BerkiGEM2007Present6">Next Section: Human Practices&gt;&gt;</a></p>
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<h1 align="center">Freeze-Drying Device</h1>
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<p align="justify">Bactoblood needs the ability to be stored for long periods in a  freeze-dried form so that it can be stockpiled and easily transported.  We therefore sought a genetic device that could improve the  Bactoblood's structural integrity during dessication. We designed two  distinct desiccation devices: one that utilizes the disaccharide  trehalose and one that involves the small molecule hydroxyectoine.</p>
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<h2 align="center">Trehalose</h2>
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<p align="center"><img name="" src="https://static.igem.org/mediawiki/2007/6/6c/Berk-Figure-otsBA.png" width="313" height="119" alt=""></p>
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<p align="justify"><img src="https://static.igem.org/mediawiki/2007/c/c2/Trehalosestructure.jpg" alt="" name="" width="266" height="180" align="left">The genes in the trehalose biosynthesis pathway are  pulled right out of the E.coli genome and consist of  otsA and otsB. By overexpressing these genes after bactoblood has reached saturation, the can protect itself from damage during freeze drying without having to add exogenous protectants.  OtsB encodes for a 29.1-kDa trehalose-6-phosphate  phosphatase and otsA encodes for a 53.6-kDa trehalose-6-phosphate  synthase. Together they make up a trehalose-6-phosphate  synthase/phosphatase complex. Naturally, the 3' end of the otsB coding  region overlaps the 5' end of the otsA coding region by 23 nucleotides.  The Trehalose helps the frozen dehydrated cells to recover back to a  functioning hydrated state. Not all cells survive the freeze drying  process but Trehalose increases the rate of success.</p>
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<p align="justify">&nbsp;</p>
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<h2 align="center">Hydroxyectoine</h2>
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<p align="justify">The hydroxyectoine biosynthesis pathway present in Streptomyces Chyrsomallus consists of four genes: thpA, thpB, thpC and thpD.  In our system, we plan on The other three genes are there to initiate the  process. When inserted into a bacterium, hydroxyectoine will prevent  dessication and many other extremes. It will protect the bacteria's  proteins and cell membrane.</p>
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<h2 align="center">Lyophilization</h2>
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<p align="center"><img src="https://static.igem.org/mediawiki/2007/9/93/Trehalose.jpg" alt="" name="" width="827" height="503"></p>
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<p align="center">The image above demonstrates that trehalose stabilizes the cell's lipid membrane and prevents leakage upon rehydration.</p>
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<p align="center">&nbsp; </p>
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<p align="center"><a href="https://2007.igem.org/Berkeley_UC">&lt;&lt;&lt; Return to UC Berkeley iGEM 2007 </a></p>
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<p align="center"><a href="https://2007.igem.org/BerkiGEM2007Present5">&lt;&lt;Previous Section: Growth Control</a> | <a href="https://2007.igem.org/BerkiGEM2007Present6">Next Section: Human Practices&gt;&gt;</a></p>
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<p align="center">&nbsp;</p>
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<b><a href="https://2007.igem.org/Berkeley_UC"><< Back to UC Berkeley iGEM 2007</a> <font face="Arial">•</font> <a href="https://2007.igem.org/BerkiGEM2007Present6">Next: Human Practices >></a></b><br /></html>
 
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==A Freeze-Drying Device==
 
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Bactoblood needs the ability to be stored for long periods in a freeze-dried form so that it can be stockpiled and easily transported.  We therefore sought a genetic device that could improve the Bactoblood's structural integrity during dessication.  We designed two distinct desiccation devices:  one that utilizes the disaccharide trehalose and one that involves the small molecule hydroxyectoine.
 
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===Trehalose===
 
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Trehalose is pulled right out of the E.coli genome and consists of the genes otsA and otsB. Naturally otsB comes first because the production of that is more important that otsA so in my operon otsB comes before otsA.
 
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Together the sugar Trehalose is produced which helps the cell recover from freeze-drying. OtsB encodes for a 29.1-kDa trehalose-6-phosphate phosphatase and otsA encodes for a 53.6-kDa trehalose-6-phosphate synthase. Together they make up a trehalose-6-phosphate synthase/phosphatase complex. Naturally, the 3' end of the otsB coding region overlaps the 5' end of the otsA coding region by 23 nucleotides. The Trehalose helps the frozen dehydrated cells to recover back to a functioning hydrated state. Not all cells survive the freeze drying process but Trehalose increases the rate of success. Trehalose structure:[[Image:trehalose2.jpg]][[Image:Trehalose.jpg]]
 
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===Hydroxyectoine===
 
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[[Image:Thp and stuff.png]]<br>
 
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Streptomyces Chyrsomallus consists of many genes; however, I was only interested in 4 genes and they are thpA, thpB, thpC and thpD. These 4 genes will help initiate the production of hydroxyectioine. The 4 genes should be arranged in this order: thpA-rbs-thpB-rbs-thpC-rbs-thpD. thpD is the gene that plays a crucial role in the production of hydroxyectoine. The other three genes are there to initiate the process. When inserted into a bacterium, hydroxyectoine will prevent dessication and many other extremes. It will protect the bacteria's proteins and cell membrane.
 

Revision as of 22:47, 26 October 2007

Untitled Document

<<< Return to UC Berkeley iGEM 2007

<<Previous Section: Growth Control | Next Section: Human Practices>>

Freeze-Drying Device

Bactoblood needs the ability to be stored for long periods in a freeze-dried form so that it can be stockpiled and easily transported. We therefore sought a genetic device that could improve the Bactoblood's structural integrity during dessication. We designed two distinct desiccation devices: one that utilizes the disaccharide trehalose and one that involves the small molecule hydroxyectoine.

Trehalose

The genes in the trehalose biosynthesis pathway are pulled right out of the E.coli genome and consist of otsA and otsB. By overexpressing these genes after bactoblood has reached saturation, the can protect itself from damage during freeze drying without having to add exogenous protectants. OtsB encodes for a 29.1-kDa trehalose-6-phosphate phosphatase and otsA encodes for a 53.6-kDa trehalose-6-phosphate synthase. Together they make up a trehalose-6-phosphate synthase/phosphatase complex. Naturally, the 3' end of the otsB coding region overlaps the 5' end of the otsA coding region by 23 nucleotides. The Trehalose helps the frozen dehydrated cells to recover back to a functioning hydrated state. Not all cells survive the freeze drying process but Trehalose increases the rate of success.

 

Hydroxyectoine

The hydroxyectoine biosynthesis pathway present in Streptomyces Chyrsomallus consists of four genes: thpA, thpB, thpC and thpD. In our system, we plan on The other three genes are there to initiate the process. When inserted into a bacterium, hydroxyectoine will prevent dessication and many other extremes. It will protect the bacteria's proteins and cell membrane.

Lyophilization

The image above demonstrates that trehalose stabilizes the cell's lipid membrane and prevents leakage upon rehydration.

 

<<< Return to UC Berkeley iGEM 2007

<<Previous Section: Growth Control | Next Section: Human Practices>>

 

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