Mississippi State University 2006
From 2006.igem.org
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* Our purpose for iGEM 2006 is to build a machine containing a reporter capable of detecting the presence of H<sub>2</sub> and producing YFP if H<sub>2</sub> is present. | * Our purpose for iGEM 2006 is to build a machine containing a reporter capable of detecting the presence of H<sub>2</sub> and producing YFP if H<sub>2</sub> is present. | ||
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+ | *Our team chose to test the hybB promoter to see if it had hydrogen sensing capabilities. We also placed switches in the constructions in order to be able to control our detectors. For a reporter, we chose a YFP. | ||
* We built the following two constructs:<br> | * We built the following two constructs:<br> | ||
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<h3>Results</h3> | <h3>Results</h3> | ||
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+ | :[[Image:MSUResultsErrorbars.JPG]] | ||
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+ | * Part BBa_J43000 | ||
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+ | * No Hydrogen, No tetR = fluorescence | ||
+ | * No Hydrogen, with tetR = Repressed Transcription at R0040 tetR, no fluorescence (except for some leakage). | ||
+ | * Hydrogen + tetR = start trqanscription at hybB, produce tetR, increased repression of operon at R0040 tetR. no fluorescence. | ||
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+ | * Part BBa_J43001 | ||
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+ | * No Hydrogen, No inducer = no fluorescence (except for a bit of leakage possibly). | ||
+ | * No Hydrogen, Inducer = Start Transcription at lacZ+PL, fluoresce. | ||
+ | * Hydrogen + Inducer = start transcription at hybB, produce lacI, competitive inhibition of operon at lacZ+PL. Less fluorescence transcribed. | ||
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+ | <h3>Discussion</h3> | ||
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+ | * Complications: | ||
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+ | * BBa_J43000 was a failed part, we learned during completion of the construction and testing that there was a flaw in the design of the composite part. Although the hybB promoter does appear to detect hydrogen, the YFP is inhibited with the addition of tetR. Therefore, we cannot detect a change in fluorescence when hydrogen is added. | ||
+ | * For the BBa_J43001, calculated error provides a possible explaination for why 100% hydrogen concentration fluoresced more than 75% hydrogen concentration. More testing required to determine the 100% hydrogen result. | ||
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+ | * Achievements: | ||
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+ | * The results of our test agree with possible success for part BBa_J43001. More testing must be performed to further prove our part as a hydrogen detector. | ||
+ | * Our machine confirms the success of hybB, BBa_Q04121, and BBa_E0430 as a composite part | ||
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<h3>Accomplishments</h3> | <h3>Accomplishments</h3> | ||
+ | * October 25- Gather results from final tests and analyze. | ||
+ | * October 24- Took pictures of vials for final tetsing. | ||
+ | * October 23- Performed gassing for final testing (Solid Media). | ||
+ | * October 22- Inoculation of E.coli + plasmid into solid media. | ||
+ | * October 17- Sent two constructions to the registry, and registered our parts on the website. | ||
+ | * October 5- Analyzed results of second test. | ||
+ | * October 4- Took pictures of vials for second testing. | ||
+ | * October 3- Performed gassing for second testing (Liquid Media). | ||
+ | * October 2- Inoculation of E.coli + plasmid into liquid media. | ||
+ | * September 29- Built testing station to be used with spectrometer. | ||
+ | * September 22- Analyzed results of first test. | ||
+ | * September 21- Took pictures of vials in first test. | ||
+ | * September 20- Performed gassing of first test (Solid Media). | ||
+ | * September 18- Inoculation of E.coli + plasmid into solid media. | ||
+ | * September 12-14- Gathered resources to begin testing. | ||
+ | * August 29-September 8- Searched for the instrumentation needed to do the testing. | ||
+ | * August 24- Digested plasmids to test the ligation of HybB (successful). | ||
+ | * August 22- Sequenced promoter for verification | ||
* August 18- Group meeting with faculty, Publicity and preparation for Jamboree discussed. | * August 18- Group meeting with faculty, Publicity and preparation for Jamboree discussed. | ||
* August 16- Planning for Jamboree with help from James. | * August 16- Planning for Jamboree with help from James. |
Latest revision as of 19:33, 30 October 2006
The TeamFaculty Members:
Students:
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Introduction
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H2 Reporter
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Results
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Discussion
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Accomplishments
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