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

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Dec 10, 2022 374 likes •537 views

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

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1.Motivation and Background

2. Lab Work
3. Policy and Practice

SLIDE 3

Sterility
Toxicity
Acidity

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

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1.50E+09 2.00E+09 2.50E+09 3.00E+09 3.50E+09 4.00E+09 0.5 1 1.5 2 2.5 3 3.5 4 Number of cells Hours after acid shock

2.00E+08
1.00E+08

0.00E+00 1.00E+08 2.00E+08 Growth rate (cells/hour)

pH 3 pH 4 pH 5 pH 7

pH 5 pH 7 pH 4 pH 3 Effects of pH on the growth rate of T7 Expression cells. Time points taken every 30

minutes. Cells were

resuspended in varying pH LB’s to be acid shocked. Cells were grown at 37˚C at 250 rpm, n=2 average shown. Top- Number of cells from

ptical density.

Bottom-growth rates of cells

ver time.

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1.00E+09 1.50E+09 2.00E+09 2.50E+09 3.00E+09 0.5 1 1.5 2 2.5 3 3.5 4 Number of cells Hours after acid shock

60 and 30 min Uninduced 15 min 0 min

1.20E+08
1.00E+08
8.00E+07
6.00E+07
4.00E+07
2.00E+07

0.00E+00 Growth Rate (cells/hour)

Uninduced 60 min 30 min 15 min 0 min

Acid response of CFA mutants . Time points taken every 30 minutes. Cells were resuspended in a LB pH of 3 to be acid

shocked. CFA was induced with 1 mM

IPTG at indicated times before shock. Cells were grown at 37˚C at 250 rpm, n=3 average shown. Top- Number of cells from optical density. Bottom-growth rates of cells over time.

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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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Akshay Sethi Ambercycle iGEM 2012: UC Davis Freedanz Ferdinand Morph Bio iGEM 2012: University College of London David Brown Mycodev iGEM 2012: University of Alberta Upcycled Aromatics

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University of Michigan Software Strategies for Resolving and Avoiding IP Conflicts Bielefield University Best Practices for Reducing the Risk Associated with Dislcosing Dual Use Technologies

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Intellectual Property Law Investment Group

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