View source for Team:Freiburg/Project/pRIG15 18


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<h1 class="sectionedit1">pRIG15_18</h1>
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<em>Treponama pallidum</em> is responsible for the sexually transmitted disease Syphillis with over 12 million new cases each year worldwide. <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup>
For our project we used the sequence of the bacterioferritin protein TpF1, an antigen of <em>Treponema pallidum</em> which reacts specifically with infected human serum. <sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup> Additionally this antigen shows strong antibody responses what makes it good target for antigen-antibody interaction studies.<sup><a class="fn_top" href="#fn__3" id="fnt__3" name="fnt__3">3)</a></sup>
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<a class="media" href="https://static.igem.org/mediawiki/2015/6/60/Freiburg_labjournal-cloning-prig15_18.jpg" title="labjournal:cloning:prig15_18.jpg"><img alt="" class="mediacenter" src="https://static.igem.org/mediawiki/2015/6/60/Freiburg_labjournal-cloning-prig15_18.jpg" width="500"/></a>
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To insert the sequence for TpF1 into pSB1C3 we designed Gibson primers with compatible overhangs that also included the start codon ATG. This fragment was amplified via PCR (Link zum Labjournal-Eintrag) and then assembled with the digested pSB1C3 backbone using Gibson assembly.
To prove correct insertion of our fragment we did a test digest (Link Labjournal) and sent the whole plasmid for sequencing.
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<div class="fn"><sup><a class="fn_bot" href="#fnt__1" id="fn__1" name="fn__1">1)</a></sup>
<a class="urlextern" href="http://www.ncbi.nlm.nih.gov/pubmed/18510892" rel="nofollow" target="_Blank" title="http://www.ncbi.nlm.nih.gov/pubmed/18510892">Daskalakis 2008 Syphilis: continuing public health and diagnostic challenges. Current HIV/AIDS reports</a></div>
<div class="fn"><sup><a class="fn_bot" href="#fnt__2" id="fn__2" name="fn__2">2)</a></sup>
<a class="urlextern" href="http://www.ncbi.nlm.nih.gov/pubmed/20385758" rel="nofollow" target="_Blank" title="http://www.ncbi.nlm.nih.gov/pubmed/20385758">McGill et al. 2010. Characterization and serologic analysis of the Treponema pallidum proteome. Infection and immunity </a></div>
<div class="fn"><sup><a class="fn_bot" href="#fnt__3" id="fn__3" name="fn__3">3)</a></sup>
<a class="urlextern" href="http://www.ncbi.nlm.nih.gov/pubmed/23945159" rel="nofollow" target="_Blank" title="http://www.ncbi.nlm.nih.gov/pubmed/23945159">Jiang et al. 2013 Evaluation of the recombinant protein TpF1 of Treponema pallidum for serodiagnosis of syphilis. Clinical and vaccine immunology</a></div>
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