Team:Prairie View/Project

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''Prairie View Team is currently developing a molecular biosensor that will detect bivalent metals at different concetrations. The data collected from testing our biosensor will be incorporated with an Electronic-Nose(E-Nose)in attempt to gain specificity within the sensor. ''
 
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Our work is aimed at developing a biosensor for the detection of cations by introducing electron network system proteins into the chassy best suited to receive an enhanced energy flow. The flow of electrons is fueled through the network of oxidation reactions of bivalent metals across Ion channels and Cytochrome complexes. By facilitating the flow of electrons in a cell, we have showed an increase of ATP production. Bacterial cells use ATP for anabolic reactions, including the synthesis of DNA. Through our electron network system, we can enhance the expression of DNA, increasing cell replication along with the fluorescence of our reporters. Together, these incorporated proteins enhance the capacity of our sensor device.<br><br>
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== '''Overall project''' ==
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Our goal is to establish specificity within the sensing device. To accomplish this, we have selected new vectors (BSK and pET) and have designed primers to replicate the parts used in previous experiments along with the addition of cytochrome C. Our approach in designing these parts is to assemble ligations with a more modular construction enabling specificity in our sensor. We can now test with each metal alone and in combinations with and without Cytochrome C & Ion Channels.<br><br>
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Theobjective of the project is to assemble different parts for the construction of a molecular DNA device to specifically detect different metal ions as well as their concentrations. The metals being tested include but are not limited to vanadium, nickel, and iron. Our approach to gain specificity is to use various computational models to design a predictable biological system. This biological system will be used to specify the particular metal, as well as its concentration. This can be applied to plant and animal systems.
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Our testing system is designed for training a computer model with the data on an accept/reject basis. The targeted divalent cations are accepted by the model because their electrical configurations allow oxidation for the electron network system. Introduced anions will be used for rejected data because their valence decline oxidation and consequently will not produce the ATP/fluorescent values that the model is trained to accept. This accept/reject basis enhances the degree of specificity of our sensor and of the computational model.<br><br>
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== Project Details==
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To expand on this research, we would like to design our own proteins; new ion channels and cytochrome complexes that are specific to the electrical configurations of the target ions. These electrically gated proteins would discriminate passage based on the energy of the ion. This kind of project would involve a collaborative field effort and we are seeking cooperative interest.
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=== Assembly ===<br>
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We have designed our own primers to replicate and amplify parts that were previously used along with some new proteins that have been synthesized. Our primer design includes compatible restriction sites flanking each part which will enable our team to modularly assemble the parts.
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!align="center"|[[Team:Prairie_View|Home]]
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The primer design seemed to have worked better with some parts more than others and multiple PCR attempts have been carried out in order to amplify each part as specified. Our RBS was synthesized separately as a Forward and Reverse oligos surrounded by compatible flanking restriction sites. The complimentary strands were annealed together, digested and gel purified.
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!align="center"|[[Team:Prairie_View/Team|The Team]]
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!align="center"|[[Team:Prairie_View/Project|The Project]]
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Ligations were complicated by this two step reaction of connecting an RBS to each part. New primers have been designed with RBS incorporated into each part.
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!align="center"|[[Team:Prairie_View/Parts|Parts Submitted to the Registry]]
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!align="center"|[[Team:Prairie_View/Modeling|Modeling]]
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=== The Experiments ===
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!align="center"|[[Team:Prairie_View/Notebook|Notebook]]
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=== Part 3 ===
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== Results ==
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Latest revision as of 19:14, 29 October 2008

Prairie View iGEM.jpg
Home The Team The Project Parts Submitted to the Registry Modeling Notebook



Our work is aimed at developing a biosensor for the detection of cations by introducing electron network system proteins into the chassy best suited to receive an enhanced energy flow. The flow of electrons is fueled through the network of oxidation reactions of bivalent metals across Ion channels and Cytochrome complexes. By facilitating the flow of electrons in a cell, we have showed an increase of ATP production. Bacterial cells use ATP for anabolic reactions, including the synthesis of DNA. Through our electron network system, we can enhance the expression of DNA, increasing cell replication along with the fluorescence of our reporters. Together, these incorporated proteins enhance the capacity of our sensor device.

Our goal is to establish specificity within the sensing device. To accomplish this, we have selected new vectors (BSK and pET) and have designed primers to replicate the parts used in previous experiments along with the addition of cytochrome C. Our approach in designing these parts is to assemble ligations with a more modular construction enabling specificity in our sensor. We can now test with each metal alone and in combinations with and without Cytochrome C & Ion Channels.

Our testing system is designed for training a computer model with the data on an accept/reject basis. The targeted divalent cations are accepted by the model because their electrical configurations allow oxidation for the electron network system. Introduced anions will be used for rejected data because their valence decline oxidation and consequently will not produce the ATP/fluorescent values that the model is trained to accept. This accept/reject basis enhances the degree of specificity of our sensor and of the computational model.

To expand on this research, we would like to design our own proteins; new ion channels and cytochrome complexes that are specific to the electrical configurations of the target ions. These electrically gated proteins would discriminate passage based on the energy of the ion. This kind of project would involve a collaborative field effort and we are seeking cooperative interest.




Home The Team The Project Parts Submitted to the Registry Modeling Notebook