Designed by: Katie Knister   Group: iGEM12_Caltech   (2012-10-01)

Bacterial Animation

For the results of the bacterial animation project, we have successfully created two constructs using an mCherry plasmid obtained from the Murray Lab at Caltech that we have submitted to the Parts Registry as BioBricks: mCherry-LVA in pSB1C3 and mCherry-AAV in pSB1C3. In parallel we inserted mCherry into R0040. The sequencing data showed all constructs have been built successfully. We ran a RFP assay on our parts in R0040 (with a tetR promoter) and found that our constructs behaved exactly as expected. We ran a characterization assay measuring the fluorescence of R0040, mCherry-LVA, mCherry-AAV, and mCherry (no degradation tag) both with and without aTc, which binds to the tetR repressor. As we hypothesized, the untagged cells produced over 50 times more fluorescent protein than those with degradation tags. The mCherry-LVA/AAV still had more fluorescence than R0040, showing that the protein was produced successfully and the degradation tags worked. Also as we predicted, the LVA had less fluorescence than the AAV, demonstrating that LVA degrades more quickly than AAV.

The picture below shows the same data from the previous graph without the untagged.

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