Difference between revisions of "Team:Freiburg/Project/pRIG15 7"

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The Varicella Zoster Virus also called Human Herpes Virus Type 3 causes chickenpox after first infection and remains as a latent infection. The sequence we used during our project codes for the extracellular part of the glycoprotein E <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup>, which is the most abundant glycoprotein on the surface of infected cells. <sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup>
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The Varicella Zoster Virus, also called Human Herpes Virus Type 3, causes <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Diseases#Zoster" title="Varicella">chickenpox</a> after first infection and remains as a latent infection. The sequence we used during our project codes for the extracellular part of the glycoprotein E <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup>, which is the most abundant glycoprotein on the surface of infected cells. <sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup>
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<a class="media" href="https://static.igem.org/mediawiki/2015/1/15/Freiburg_labjournal-cloning-prig15_7.jpg" title="labjournal:cloning:prig15_7.jpg"><img alt="" class="mediacenter" src="https://static.igem.org/mediawiki/2015/1/15/Freiburg_labjournal-cloning-prig15_7.jpg" width="500"/></a>
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                      <p><strong>Figure 1: pRIG15_7.</strong> BBa_K1621001 inserted into the submission backbone pSB1C3.</p>
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To insert the sequence for Varicella Zoster glycoprotein E 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.
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To insert the sequence for Varicella Zoster glycoprotein E into <a class="urlextern" href="http://parts.igem.org/Part:pSB1C3" target="_blank" title="pSB1C3">pSB1C3</a> 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#PCR5_7_9_11" title="PCR5_7_9_11">PCR</a> and then assembled with the digested <a class="urlextern" href="http://parts.igem.org/Part:pSB1C3" target="_blank" title="pSB1C3">pSB1C3</a> backbone using <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Classic_vs_Gibson" title="gibson">Gibson</a> assembly.
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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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To prove correct insertion of our fragment we did a <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Labjournals/Cloning/August#TDbb6_7" title="TDbb6_7">test digest</a> and sent the whole plasmid for sequencing.
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Link to GeneBank file: <a class="media" href="https://static.igem.org/mediawiki/2015/3/34/Freiburg_2015_BBa_K1621001.gb" title="2015_Freiburg_BBa_K1621001" src="https://static.igem.org/mediawiki/2015/3/34/Freiburg_2015_BBa_K1621001.gb">BBa_K1621001.gb</a>.
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Link to Registry: <a href="http://parts.igem.org/Part:BBa_K1621001" target="_blank">BBa_K1621001</a>
 
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Latest revision as of 22:32, 16 September 2015

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pRIG15_7

The Varicella Zoster Virus, also called Human Herpes Virus Type 3, causes chickenpox after first infection and remains as a latent infection. The sequence we used during our project codes for the extracellular part of the glycoprotein E 1), which is the most abundant glycoprotein on the surface of infected cells. 2)

Figure 1: pRIG15_7. BBa_K1621001 inserted into the submission backbone pSB1C3.

To insert the sequence for Varicella Zoster glycoprotein E 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 our fragment we did a test digest and sent the whole plasmid for sequencing.

Link to GeneBank file: BBa_K1621001.gb.
Link to Registry: BBa_K1621001

1) Thomsson et al. 2011. Recombinant glycoprotein E produced in mammalian cells in large-scale as an antigen for varicella-zoster-virus serology. Journal of virological methods
2) Mo et al. 2003. Varicella-zoster virus infection facilitates VZV glycoprotein E trafficking to the membrane surface of melanoma cells. Journal of medical virology