Difference between revisions of "Team:Freiburg/Project/pRIG15 18"
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| − | < | + | <i>Treponama pallidum</i> is responsible for the sexually transmitted disease <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Diseases#Syphilis" title="Syphilis">Syphillis</a> 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 < | + | For our project we used the sequence of the bacterioferritin protein TpF1, an antigen of <i>Treponema pallidum</i> 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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| − | <div class=" | + | <div class="image_box right"> |
| − | <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=" | + | <div class="thumb2 trien" style="width:300px"><div class="thumbinner"> |
| + | <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="300"/></a></div><strong>Figure 1: pRIG15_18.</strong> BBa_K621005 inserted into the submission vector pSB1C3.</div> | ||
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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 <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Labjournals/Cloning/August#PCR6-18" title="PCR6-18">PCR</a> and then assembled with the digested pSB1C3 backbone using Gibson | + | 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 <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Labjournals/Cloning/August#PCR6-18" title="PCR6-18">PCR</a> and then assembled with the digested pSB1C3 backbone using Gibson Assembly. |
| − | To prove correct insertion of | + | To prove correct insertion of the fragment we performed a <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Labjournals/Cloning/August#TD8_10_18" title="TD8_10_18">test digest</a> and verified the part by sequencing. |
</br> | </br> | ||
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| − | + | Get the sequence in genebank format: <a class="media" href="https://static.igem.org/mediawiki/2015/b/b0/Freiburg_2015_BBa_K1621005.gb" title="2015_Freiburg_BBa_K1621005" src="https://static.igem.org/mediawiki/2015/b/b0/Freiburg_2015_BBa_K1621005.gb">BBa_K1621005.gb</a>. | |
</p> | </p> | ||
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<div class="footnotes"> | <div class="footnotes"> | ||
| + | <br> | ||
<div class="fn"><sup><a class="fn_bot" href="#fnt__1" id="fn__1" name="fn__1">1)</a></sup> | <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> | <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> | ||
Revision as of 12:50, 14 September 2015
pRIG15_18
Treponama pallidum is responsible for the sexually transmitted disease Syphillis with over 12 million new cases each year worldwide. 1) For our project we used the sequence of the bacterioferritin protein TpF1, an antigen of Treponema pallidum which reacts specifically with infected human serum. 2) Additionally, this antigen shows strong antibody responses what makes it good target for antigen-antibody interaction studies.3)
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 and then assembled with the digested pSB1C3 backbone using Gibson Assembly. To prove correct insertion of the fragment we performed a test digest and verified the part by sequencing. Get the sequence in genebank format: BBa_K1621005.gb.
