Difference between revisions of "Team:Freiburg/Project/pRIG15 8"

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Infection with the Herpes Simplex Virus 1 leads to life-long persistence of the virus inside the body as a latent infection. <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup> The glycoprotein G is one of 11 glycoproteins expressed by HSV1. <sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup> The sequence we used for our experiment was obtained from
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Infection with the <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Diseases#Herpes" title="Herpes simplex">Herpes Simplex Virus 1</a>
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leads to life-long persistence of the virus inside the body as a latent infection. <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup> The glycoprotein G is one of 11 glycoproteins expressed by HSV-1. <sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup> The sequence we used for our experiment was obtained from
 
Jaaskelainen et al., 2009. <sup><a class="fn_top" href="#fn__3" id="fnt__3" name="fnt__3">3)</a></sup>
 
Jaaskelainen et al., 2009. <sup><a class="fn_top" href="#fn__3" id="fnt__3" name="fnt__3">3)</a></sup>
 
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                      <p><strong>Figure 1: pRIG15_8.</strong> BBa_K1621002 inserted into the submission backbone pSB1C3.</p>
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To insert the sequence for HSV-1 glycoprotein G 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#PCR6-18" title="PCR6-18">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 <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Labjournals/Cloning/August#TD8_10_18" title="TD8_10_18">test digest</a> and sent the whole plasmid for sequencing.
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To insert the sequence for HSV1 glycoprotein G 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 prove correct insertion of our fragment we did a test digest (Link Labjournal) and sent the whole plasmid for sequencing.
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Link to GeneBank file: <a class="media" href="https://static.igem.org/mediawiki/2015/2/22/Freiburg_2015_BBa_K1621002.gb" title="2015_Freiburg_BBa_K1621002" src="https://static.igem.org/mediawiki/2015/2/22/Freiburg_2015_BBa_K1621002.gb">BBa_K1621002.gb</a>.
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Link to Registry: <a href="http://parts.igem.org/Part:BBa_K1621002" target="_blank">BBa_K1621002</a>
 
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Latest revision as of 22:28, 16 September 2015

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pRIG15_8

Infection with the Herpes Simplex Virus 1 leads to life-long persistence of the virus inside the body as a latent infection. 1) The glycoprotein G is one of 11 glycoproteins expressed by HSV-1. 2) The sequence we used for our experiment was obtained from Jaaskelainen et al., 2009. 3)

Figure 1: pRIG15_8. BBa_K1621002 inserted into the submission backbone pSB1C3.

To insert the sequence for HSV-1 glycoprotein G 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_K1621002.gb.
Link to Registry: BBa_K1621002

1) Corey et al. 1994. The current trend in genital herpes. Progress in prevention. Sexually transmitted diseases
2) Rekabdar et al. 2002. Dichotomy of glycoprotein g gene in herpes simplex virus type 1 isolates. Journal of clinical microbiology
3) Jaaskelainen et al. 2009. Serological microarray for detection of HSV-1, HSV-2, VZV, and CMV antibodies. Journal of virological methods.