Difference between revisions of "Team:Freiburg/Project/pRIG15 8"
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| − | 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 <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 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. |
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. | 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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Latest revision as of 22:28, 16 September 2015
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)
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
