Team:Missouri Miners/Project

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(Project Details)
(Project Details)
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Some of the techniques that have been used thus far in pursuing this project have been:
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Laboratory techniques that have been utilized by team members working on the project:
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#PCR amplification of the MacA and other cytocromes using PCR primers that added standard prefix and suffix sequences
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#Cloning the cytocrome PCR products into a commercial cloning vector
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#PCR amplification of the MacA and other cytochromes using PCR primers that added standard prefix and suffix sequences
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#Digesting the cytocrome plasmids to obtain the promoter genes of interest
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#Cloning the cytochrome PCR products into a commercial cloning vector
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#Digesting the cytochrome plasmids to obtain the promoter genes of interest
#Separating and isolating the enzyme digest fragments through gel electrophoresis
#Separating and isolating the enzyme digest fragments through gel electrophoresis
#Ligating the individual cytochrome promoters together
#Ligating the individual cytochrome promoters together

Revision as of 08:30, 4 August 2009

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PROJECT - Microbial Fuel Cell:

The goal of this research is the manipulation of E.Coli; granting them the ability to release electrons in an aerobic enviroment. This project utilizes geobacter's cytocromes, which makes it's extracellular electron transfer possible. The cytocromes OmcB,OmcE,OmcS, and MacA are our priority since they are the major electron smugglers out of the cell. By isolating and placing these cytocromes, some of which are naturally occur in E.Coli,into bricks we hope to make the electical application possible in other bacterial species.


Overall project

Optimization of electron shuffle to external surfaces such as anodes is a primary goal. Geobacter sulfurreducens happens to be our model bacteria due to its ability in nature to efficiently export electrons extracelluarly. E. coli can be the chassis for this experiment due to its genome already containing some key proteins in our preferred pathway. The proteins, such as extracellular pilin, MacA, and many other cytochromes, which E. coli does not have will be isolated from Geobacter sulfurreducens and introduced into E. coli to formulate the most optimal pathway for generating electronmotive force in a microbial fuel cell apparatus.

Some problems will be faced concerning plasmid engineering and the simple fact that Geobacter is anaerobic and E. coli is aerobic. Also, the role of Geobacter's pili in extracellular electron transfer is not clearly understood and could create a significant problem since e. coli does not have such pili. As a team, we will push in the right direction harder than an emf on the internal resistivity of a toroid. Many diverse team members will work in concert utilizing Missouri S&T’s dominating Electrical, Chemical, and Biological Engineering undergraduates along with Biological Science masterminds.

Project Details

Procedure:

Laboratory techniques that have been utilized by team members working on the project:

  1. PCR amplification of the MacA and other cytochromes using PCR primers that added standard prefix and suffix sequences
  2. Cloning the cytochrome PCR products into a commercial cloning vector
  3. Digesting the cytochrome plasmids to obtain the promoter genes of interest
  4. Separating and isolating the enzyme digest fragments through gel electrophoresis
  5. Ligating the individual cytochrome promoters together
  6. Transforming bacterial cells with the recombined cytochrome plasmids
Extracellular Electron Transport Chain


Possible Applications: The applications of a interchangeable electron transfer gene part are limitless. The cytochrome cocktail could be used as a signal within bacteria. It would produce small currents which would be utilized in a similar fashion to fluorescent proteins as markers and indicators of cellular activity. Micro and nano-electronics could be adapted to use electron transfer modified bacteria as a cheap and effective power source. Medical research could benefit by using modified bacteria in areas of the body such as the stomach and intestines. The bacteria could be developed to act as a long term mobile monitor inside the human body.