Team:Wisconsin-Madison/results

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(Stationary Phase Induction)
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===Background===
===Background===
'''gadAp: A pH Sensitive Promoter'''
'''gadAp: A pH Sensitive Promoter'''
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Based on available literature (cite), 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). In order to test this, we combined the gadA promoter with a RFP gene (BBa_K318513). This part
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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).
===Low pH Induction===
===Low pH Induction===

Revision as of 17:33, 27 October 2010

Encapsulation

Parts used in Encapsulation Experiments
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

Cell Survivability Testing Graphs.jpg

Conclusion


Timed Lysis

Parts used in these Experiments
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).

Low pH Induction

See procedure at experimental protocols page.

Stationary Phase Induction

See procedure at experimental protocols page.

Conclusion