Difference between revisions of "Team:BostonU/App 1/Results"

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<h3>Results</h3>
 
<h3>Results</h3>
 +
 +
<p>To test this system, different reporters were used for each split protein. The reporters contain a
 +
constitutive promoter succeeded by an iRFP coding sequence, then succeeded by an inverted mRuby
 +
coding sequence. These coding sequences are flanked by the recombination sites associated with each
 +
integrase or RDF. In the case of the RDF splits, the full integrase is co-transfected. If the split protein is
 +
active in the presence of the inducer it will invert the coding sequences turning off the expression of
 +
iRFP, and turning on expression of mRuby. The fluorescence levels at the wavelengths associated with
 +
each of these proteins is assayed. By measuring the fluorescence of mRuby we can confirm or reject the
 +
viability of the split integrase or RDF.</p>
 +
 +
<p>When carrying out this experiment, each split integrase and RDF were tested along with a
 +
negative control that contained the split integrase in the absence of the inducer. A positive control
 +
which contained the full integrase or RDF was tested. By comparing the split protein activity to the
 +
negative and positive control, we can assay induced activity vs basal activity and compare these
 +
activities to the full wild type protein.</p>
  
 
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Revision as of 04:45, 15 September 2015






Motivation Design Results

Results

To test this system, different reporters were used for each split protein. The reporters contain a constitutive promoter succeeded by an iRFP coding sequence, then succeeded by an inverted mRuby coding sequence. These coding sequences are flanked by the recombination sites associated with each integrase or RDF. In the case of the RDF splits, the full integrase is co-transfected. If the split protein is active in the presence of the inducer it will invert the coding sequences turning off the expression of iRFP, and turning on expression of mRuby. The fluorescence levels at the wavelengths associated with each of these proteins is assayed. By measuring the fluorescence of mRuby we can confirm or reject the viability of the split integrase or RDF.

When carrying out this experiment, each split integrase and RDF were tested along with a negative control that contained the split integrase in the absence of the inducer. A positive control which contained the full integrase or RDF was tested. By comparing the split protein activity to the negative and positive control, we can assay induced activity vs basal activity and compare these activities to the full wild type protein.