1.Motivation and Background 2. Lab Work 3. Policy and Practice - PowerPoint PPT Presentation

1.Motivation and Background 2. Lab Work 3. Policy and Practice

• Sterility • Toxicity • Acidity

• CFA: Cyclopropane Fatty Acid • UFA: Unsaturated Fatty Acid • SAM: S-Adenosyl Methionine

4.00E+09 3.50E+09 pH 5 Number of cells pH 7 3.00E+09 pH 4 2.50E+09 2.00E+09 pH 3 1.50E+09 pH 5 pH 7 2.00E+08 0 0.5 1 1.5 2 2.5 3 3.5 4 Hours after acid shock Effects of pH on the growth pH 4 1.00E+08 rate of T7 Expression cells. Growth rate (cells/hour) Time points taken every 30 minutes. Cells were resuspended in varying pH 0.00E+00 LB’s to be acid shocked. Cells were grown at 37˚C at 250 rpm, n=2 average shown. -1.00E+08 Top- Number of cells from pH 3 optical density. Bottom-growth rates of cells -2.00E+08 over time.

3.00E+09 2.50E+09 60 and 30 min Number of cells Uninduced 2.00E+09 15 min 0 min 1.50E+09 0.00E+00 1.00E+09 0 0.5 1 1.5 2 2.5 3 3.5 4 -2.00E+07 Hours after acid shock Growth Rate (cells/hour) Acid response of CFA mutants . Time 60 min 30 min -4.00E+07 points taken every 30 minutes. Cells were Uninduced resuspended in a LB pH of 3 to be acid -6.00E+07 shocked. CFA was induced with 1 mM IPTG at indicated times before shock. -8.00E+07 Cells were grown at 37˚C at 250 rpm, n=3 15 min average shown. Top- Number of cells from optical -1.00E+08 0 min density. Bottom-growth rates of cells over time. -1.20E+08

• Correlation between increased acidity and decreased number of cells • With ample induction time there is a decrease in cell lysis compared to our uninduced control • This suggests CFA is responsible for this shift • Ensure CFA is being expressed • Alter pH range • Sample more frequent time points • Test the response to alkali conditions • Assess vitality of cells • Run a prolonged culture • Scaled up

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