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Team:Wisconsin-Madison/results
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===Background=== | ===Background=== | ||
'''gadAp: A pH Sensitive Promoter''' | '''gadAp: A pH Sensitive Promoter''' | ||
| - | Based on availble literature (see Project Page), we expected an increase in expression from gadAp (BBa_K318512) under both acidic pH conditions (from pH 5 to 3) and in the stationary phase (after an OD of 1 or 2). As most previous studies of gadAp had focused on gadA mRNA production, we wanted to instead explore the viability of gadAp for use in expression of protein. Given the stressful conditions under which it acts and its complex regulation, the possiblity that mRNA-level regulation or an inability of acid-stressed cells to produce large quantities of protein may exist could not be discounted. As such, we combined the gadA promoter with a RFP gene (BBa_K318513). | + | Based on availble literature (see Project Page), we expected an increase in expression from gadAp (BBa_K318512) under both acidic pH conditions (from pH 5 to 3) and in the stationary phase (after an OD of 1 or 2). As most previous studies of gadAp had focused on gadA mRNA production, we wanted to instead explore the viability of gadAp for use in expression of protein. Given the stressful conditions under which it acts and its complex regulation, the possiblity that mRNA-level regulation or an inability of acid-stressed cells to produce large quantities of protein may exist could not be discounted. As such, we combined the gadA promoter with a RFP gene (BBa_K318513) to directly test its ability to produce protein under stationary phase and acid stress conditions. |
===Low pH Induction=== | ===Low pH Induction=== | ||
Revision as of 17:34, 27 October 2010
Encapsulation
| Part Number | Function | Expression Type | Zip File |
| <partinfo>BBa_k318500</partinfo> | Produces Trascription Factor RcsB | Inducible - IPTG | 500 |
| <partinfo>BBa_k318501</partinfo> | Produces Trascription Factor RcsA | Inducible - IPTG | 501 |
| <partinfo>BBa_k318502</partinfo> | Produces Trascription Factor RcsA & RcsB | Inducible - IPTG | 502 |
| <partinfo>BBa_k200021</partinfo> | Empty Vector/Contol | Inducible - IPTG | NA |
Colonic Acid Quantification
Background
Colonic Acid is a polysaccharide containing a repeat unit with D-glucose, L-fucose, D-galactose, and D-glucuronate. Biological extracts often contain compounds, which under heating with H2SO4 yield brown products absorbing between 396 nm and 427 nm. Colonic acid can be estimated by measuring L-fucose content.
Download procedure here or continue to experimental protocol page
Results
Conclusion
Cell Survivability in Low pH
Background
Download procedure here or continue to experimental protocol page
Results
Conclusion
Timed Lysis
| Part Number | Function | Induction | Zip File |
| <partinfo>BBa_k318513</partinfo> | Produces RFP | stationary phase | 513 |
Background
gadAp: A pH Sensitive Promoter Based on availble literature (see Project Page), we expected an increase in expression from gadAp (BBa_K318512) under both acidic pH conditions (from pH 5 to 3) and in the stationary phase (after an OD of 1 or 2). As most previous studies of gadAp had focused on gadA mRNA production, we wanted to instead explore the viability of gadAp for use in expression of protein. Given the stressful conditions under which it acts and its complex regulation, the possiblity that mRNA-level regulation or an inability of acid-stressed cells to produce large quantities of protein may exist could not be discounted. As such, we combined the gadA promoter with a RFP gene (BBa_K318513) to directly test its ability to produce protein under stationary phase and acid stress conditions.
Low pH Induction
See procedure at experimental protocols page.
Stationary Phase Induction
See procedure at experimental protocols page.
Conclusion
