Team:Virginia Commonwealth/Internal/Project ideas

From 2009.igem.org

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I think looking at homologous proteins across temperature preferences would be a good start. Usually dense G-C regions are more thermostable in terms of T<sub>m</sub>, similar substitutions in amino acid sequences and tertiary/quarternary structure. Also, prions would be neat to look at, because they don't denature at 60°C, but require an extreme of 134°C for a while. Also, partially denatured prions can renature and become active and infections. [[User:Jalvin|Jalvin]] 20:39, 28 May 2009 (UTC)
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I think looking at homologous proteins across temperature preferences would be a good start. Usually dense G-C regions are more thermostable in terms of T<sub>m</sub>, similar substitutions in amino acid sequences and tertiary/quarternary structure. Also, prions would be neat to look at, because they don't denature at 60°C, but require an extreme of 134°C for a while. Also, partially denatured prions can renature and become active and infections. See [http://www.pnas.org/content/83/3/576.full.pdf Isolation of a thermostable enzyme variant by cloning and selection in a thermophile] [[User:Jalvin|Jalvin]] 20:39, 28 May 2009 (UTC)
==Extracellular enzyme scaffold==
==Extracellular enzyme scaffold==

Revision as of 20:44, 28 May 2009

Use this space to brainstorm project ideas. Go crazy with it. Feel free to make comments and suggestions to existing ideas. Please sign your comments by inserting "~~~~" after each so that we can keep track of who is communicating and when the information was posted.

Contents

Systematic characterization of promoters

Overview

Significant knowledge gaps remain in the functional and dynamic characterization of several gene promoters, and engineering decisions depend on detailed quantitative information.

Comments

  • George, do you have any good papers about this? Chris.m.gowen 01:08, 22 April 2009 (UTC)
    • Yeah, there is a recent paper from Endy et al. titled [http://www.jbioleng.org/content/pdf/1754-1611-3-4.pdf Measuring the activity of BioBrick promoters using an in vivo reference standard]. In addition, there is a promoter measurement kit that has been developed by Jason Kelly (Ginkgo Bioworks) that is distributed by the Registry (I believe it was used in the paper mentioned above). More information, including a protocol, is available [http://partsregistry.org/Measurement here]. GMcArthurIV 15:18, 17 May 2009 (UTC)

Development of thermostable enzymes

Overview

Bacteria and Archea occupy nearly every nook and cranny on Earth; from deep beneath Arctic ice in the absence of light, to temperatures over boiling in Yellowstone. As thermodynamics tells us, reaction rates increase proportional to temperature. Therefore, an industrial enzymatic process normally carried out at 37oC, could be run at higher temperatures for a faster turnover. Additionally, incubating microbes is very energy-intensive, and the knowledge of thermostable properties can also be applied to utilize thermophilic enzymes in meso- or even psychrophiles. This could lead to Quiagen kits that don't require any incubation, yet can proceed at the same rate.

Comments

I think looking at homologous proteins across temperature preferences would be a good start. Usually dense G-C regions are more thermostable in terms of Tm, similar substitutions in amino acid sequences and tertiary/quarternary structure. Also, prions would be neat to look at, because they don't denature at 60°C, but require an extreme of 134°C for a while. Also, partially denatured prions can renature and become active and infections. See [http://www.pnas.org/content/83/3/576.full.pdf Isolation of a thermostable enzyme variant by cloning and selection in a thermophile] Jalvin 20:39, 28 May 2009 (UTC)

Extracellular enzyme scaffold

Overview

A cell-associated protein scaffold which could accommodate various enzymes or binding proteins can create a cellular assembly line by drastically reducing inefficiencies of product and reactant diffusion and by aligning binding sites to targets. This concept is what makes the cellulosome of Clostridium thermocellum and other organisms so efficient at breaking down cellulose. See the [http://www.springerlink.com/content/785bm9d7uugugrdn/ review] by Schwarz for an overview of this system. A protein known as scaffoldin is a consistent part of each cellulosome, and provides the basic structure to which each catalytic enzyme attaches. In many cases a cohesin module can bind a wide range of proteins to the scaffoldin, and a dockerin module binds the scaffoldin to the cell surface. If we could export the scaffoldin and a dockerin module to E. coli, it may be possible to add the cohesin domain to the non-catalytic regions of desirable enzymes and create a synthetic "reaction line."

Problem areas

  • Protein folding may be quite different in E. coli, especially from thermophilic source organisms.
  • The dockerin module binds to the cell wall of the gram positive source organisms, which ecoli of course doesn't have...
  • Integrating a foreign cohesin domain into native enzymes is in the realm of protein engineering, with which I am not familiar
    • Here are some programs that may provide a way to verify protein docking [http://openwetware.org/wiki/Wikiomics:Docking_toolbox#Protein-protein_docking here]; Chris.m.gowen 01:46, 22 April 2009 (UTC)
  • Others?

Organisms with cellulosomes

Clostridium thermocellum
C. acetobutylicum

Comments

Yeast that make gluten free beer

Overview

Gluten is a protein (proteins?) in wheat that is in most breads and beers. Many people have allergies to these proteins, and so have to carefully avoid gluten-containing products. Maybe we could engineer a pathway to specifically break down gluten in the brewing process?

Comments

Using yeast may be more trouble than we want for the first year. chris

I'm not 21. Jalvin 20:08, 28 May 2009 (UTC)

Bacterial interaction

Overview

Some kind of social behavior, like quorum sensing or self assembly based on communicating only with neighboring cells would be cool. This would be a good way to take advantage of Advait's cellular automata expertise before he leaves. This would also be somthing that would require good quantitative characterization of our genes and could be combined with the promoter characterization stuff, maybe.