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Team:UEA-JIC Norwich/Nittygritty-bacteria
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| + | <p><h1>Making bacteria see.</h1></p> | ||
| + | <p>We are planning to transform the bacterial species ''Escherichia coli''. We chose ''E.coli'' because its genome has been completely sequenced, it has a high transformation frequency and is relatively simple to manipulate. As most iGEM projects have centered around ''E.coli'' most Biobricks are also optimised for expression in this species. We therefore wished to first use ''E.coli'' to test that the luminescence could be achieved and that a form of control was also possible, before we moved onto the more advanced eukaryotic species we planned to work with. We plan to ultimately transform ''E.coli'' with a new composite Biobrick. This will be formed of four existing Biobricks including Cph8, OmpR, Green Luciferase and Luciferin Recovery Enzyme.</p> | ||
| + | <p>Cph8 is a composite part formed of the Cph1 and Env Biobricks. Cph1 is a light sensing domain most sensitive to red light in the region of 660nm. When translated, light exposure inhibits the activity of the EnvZ histidine kinase domain. Thus, the kinase activity will only be active in the dark. When in this state, EnvZ phosphorylates the OmpR promoter to OmpR-P. This promoter is usually upstream of the OmpC gene. We are planning to place this promoter upstream of the luciferase and luciferase recovery enzyme Biobricks. Therefore, our cells will only luminesce in the dark!</p> | ||
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Revision as of 09:47, 1 July 2011
Making bacteria see.
We are planning to transform the bacterial species ''Escherichia coli''. We chose ''E.coli'' because its genome has been completely sequenced, it has a high transformation frequency and is relatively simple to manipulate. As most iGEM projects have centered around ''E.coli'' most Biobricks are also optimised for expression in this species. We therefore wished to first use ''E.coli'' to test that the luminescence could be achieved and that a form of control was also possible, before we moved onto the more advanced eukaryotic species we planned to work with. We plan to ultimately transform ''E.coli'' with a new composite Biobrick. This will be formed of four existing Biobricks including Cph8, OmpR, Green Luciferase and Luciferin Recovery Enzyme.
Cph8 is a composite part formed of the Cph1 and Env Biobricks. Cph1 is a light sensing domain most sensitive to red light in the region of 660nm. When translated, light exposure inhibits the activity of the EnvZ histidine kinase domain. Thus, the kinase activity will only be active in the dark. When in this state, EnvZ phosphorylates the OmpR promoter to OmpR-P. This promoter is usually upstream of the OmpC gene. We are planning to place this promoter upstream of the luciferase and luciferase recovery enzyme Biobricks. Therefore, our cells will only luminesce in the dark!
