Difference between revisions of "Part:BBa K773004:Experience"

 
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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how you used this part and how it worked out.
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<h2> Bacterial Animation </h2>
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For the results of the bacterial animation project, we have successfully created two constructs using an mCherry plasmid obtained from the <a href="http://www.cds.caltech.edu/~murray/wiki/Main_Page">Murray Lab</a> 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.
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<img src="https://static.igem.org/mediawiki/2012/5/52/MCherry_fluorescence.png">
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The picture below shows the same data from the previous graph without the untagged.
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<img src="https://static.igem.org/mediawiki/2012/d/d9/MCherry_fluorescence_zoom.png">
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===Applications of BBa_K773004===
 
===Applications of BBa_K773004===

Latest revision as of 21:21, 13 October 2012

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.


Applications of BBa_K773004

User Reviews

UNIQ4c817efb6ae35e29-partinfo-00000001-QINU UNIQ4c817efb6ae35e29-partinfo-00000002-QINU