Team:Michigan/Project
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The goals of the quorum sensing team are two-fold: first to characterize the response of ''E. coli'' to the ''C. vulgaris'' AI-2 mimic (which may be actual AI-2), and second to engineer ''E. coli'' to flocculate in response to the presence of ''C. vulgaris''. The first task will be accomplished by transforming a LuxS-mutant strain of ''E. coli'' (cannot produce its own AI-2) with an AI-2 reporter biobrick. We will then harvest supernatant from ''C. vulgaris'', which should contain AI-2 or its mimic, and apply it to the reporter strain to test its response. The second task will be accomplished by ligating a gene that causes over-expression of pili (characterized by the pili team) to the Lsr promoter, which is derepressed in response to AI-2. By transforming this part into LuxS-mutant ''E. coli'', we hope to create strain that will stick to algae and will flocculate only in the presence of ''C. vulgaris''. | The goals of the quorum sensing team are two-fold: first to characterize the response of ''E. coli'' to the ''C. vulgaris'' AI-2 mimic (which may be actual AI-2), and second to engineer ''E. coli'' to flocculate in response to the presence of ''C. vulgaris''. The first task will be accomplished by transforming a LuxS-mutant strain of ''E. coli'' (cannot produce its own AI-2) with an AI-2 reporter biobrick. We will then harvest supernatant from ''C. vulgaris'', which should contain AI-2 or its mimic, and apply it to the reporter strain to test its response. The second task will be accomplished by ligating a gene that causes over-expression of pili (characterized by the pili team) to the Lsr promoter, which is derepressed in response to AI-2. By transforming this part into LuxS-mutant ''E. coli'', we hope to create strain that will stick to algae and will flocculate only in the presence of ''C. vulgaris''. | ||
- | [[Image:QS_circuit.jpg|thumb|left|[10]]] | + | [[Image:QS_circuit.jpg|thumb|700px|left|FIG. 5. Regulatory mechanisms of the LsrR/phospho-AI-2 circuit in E. coli AI-2 uptake (modified from reference 70). The AI-2 uptake repressor |
+ | LsrR represses many genes including the lsr operon (comprised of lsrACDBFG) and the lsrRK operon. AI-2 can be imported back inside the cell via | ||
+ | LsrACDB. Imported AI-2 is processed as phospho-AI-2 via the kinase LsrK. Phospho-AI-2 has been reported to bind LsrR and relieve its repression | ||
+ | of the lsr transporter genes, triggering their expression. This in turn stimulates additional AI-2 uptake. DPD, 4,5-dihydroxy-2,3-pentanedione.[10]]] | ||
+ | <br style="clear: both" /> | ||
=== Results to Date === | === Results to Date === |
Revision as of 03:54, 27 October 2010