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Gene Expression
Transcript
- Designed a viability assay of mutant strain grown on low osmolarity medium.
- Also investigate effects of modulating expression with adding Zinc and Copper with and without an osmoprotectant.
- Validate results of viability assay using selected samples to perform qRT-PCR.
THANK YOU FOR LISTENING
Gene Expression
The Background
What We Learned
What We Used
The Results
wild-type OD graph
Luminescence Graph
OD Graph
- We used mutants that had insertion in mechanosensitive ion channel location - MscS.
- This phenotype showed increased zinc tolerance.
- Decreased in growth in zinc.
- No pronounced effect in copper.
- Generally decreased growth in betaine.
- Growth in presence of zinc is increased.
- No pronounced changed with copper.
- Cells growth in betaine is lower
- Luminescence in betaine is higher
- Could be cells growth quicker but die faster without betaine.
Average fold gene expression changes
Our Question
- Cells grown in betaine show higher viability than without betaine.
Lessons Learned
- Is the increased zinc tolerance due to regulation of gene expression or is it due to a modification in the MscS function.
- literature searches suggested the site of transposon insertion would cause structural disruption.
- Gene seems to be highly conserved with multiple copies present.
- Betaine in addition affects the osmotic environment as well as gene downregulation.
- Would've been nice to have the transposon inserted in the correct orientations.
The Experiment
Overview of Experimental Design
Important Points
- Be Careful not to contaminate RNA.
- Keep track of the contents of the plates.
- Ensure dilution/concentration calculations were correct.
by Andreas, Luki and Jonny
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