Difference between revisions of "Team:Freiburg/Project/pRIG15 15"
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| − | <em>Salmonella</em> Typhimurium is a pathogen causing gastrointestinal infections and which is particularly dangerous because it already gained resistence against multiple antibiotics. <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup> In collaboration with the group from Prof. Dr. Hust from the University of Braunschweig we were able to get an expression vector containing the sequence for a putative dihydroxyacid dehydratase and an additional vector bearing the respective antibody, anti-putative dihydroxyacid dehydratase <a class="urlextern" href="http://parts.igem.org/Part:BBa_K1621007" rel="nofollow" target="_Blank" title="http://parts.igem.org/Part:BBa_K1621007">Bba_K1621007</a>. <sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup> | + | <em>Salmonella</em> Typhimurium is a pathogen causing gastrointestinal infections and which is particularly dangerous because it already gained resistence against multiple antibiotics. <sup><a class="fn_top" href="#fn__1" id="fnt__1" name="fnt__1">1)</a></sup> In collaboration with the group from Prof. Dr. Hust from the University of Braunschweig we were able to get an expression vector containing the sequence for a putative dihydroxyacid dehydratase and an additional vector bearing the respective antibody, anti-putative dihydroxyacid dehydratase (<a class="urlextern" href="http://parts.igem.org/Part:BBa_K1621007" rel="nofollow" target="_Blank" title="http://parts.igem.org/Part:BBa_K1621007">Bba_K1621007</a>). <sup><a class="fn_top" href="#fn__2" id="fnt__2" name="fnt__2">2)</a></sup> |
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To insert the sequence for the Salmonella dehydroxyacid dehydratase 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) from the expression vectors we got from Prof. Dr. Hust and then assembled with the digested pSB1C3 backbone using Gibson assembly. | To insert the sequence for the Salmonella dehydroxyacid dehydratase 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) from the expression vectors we got from Prof. Dr. Hust 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 | + | 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/2015/2/2b/Freiburg_2015_BBa_K1621006.gb" title="2015_Freiburg_BBa_K1621006" src="https://static.igem.org/mediawiki/2015/2/2b/Freiburg_2015_BBa_K1621006.gb">BBa_K1621006.gb</a>. | ||
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Revision as of 12:47, 7 September 2015
pRIG15_15
Salmonella Typhimurium is a pathogen causing gastrointestinal infections and which is particularly dangerous because it already gained resistence against multiple antibiotics. 1) In collaboration with the group from Prof. Dr. Hust from the University of Braunschweig we were able to get an expression vector containing the sequence for a putative dihydroxyacid dehydratase and an additional vector bearing the respective antibody, anti-putative dihydroxyacid dehydratase (Bba_K1621007). 2)
To insert the sequence for the Salmonella dehydroxyacid dehydratase 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) from the expression vectors we got from Prof. Dr. Hust 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. Link to genebank file: BBa_K1621006.gb.
