http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&feed=atom&action=historyTeam:Marburg:Project:Tumor - Revision history2024-03-28T14:57:50ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=398043&oldid=prevRomanM89 at 03:33, 18 October 20142014-10-18T03:33:14Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>To address the goal of creating a CancerSURF for efficient and rapid targeting of cancer cells, we first had to synthesize a Strep-DARPin fusion construct </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>To address the goal of creating a CancerSURF for efficient and rapid targeting of cancer cells, we first had to synthesize a Strep-DARPin fusion construct </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>(<del class="diffchange diffchange-inline">StrepDARPidin </del><html><a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank"><del class="diffchange diffchange-inline">BBa_K1329000</del></a></html>). </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>(<html><a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank"><ins class="diffchange diffchange-inline">StrepDARPidin</ins></a></html>). </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Template DNA for the tetrameric <html><a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a></html> construct was commercially </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Template DNA for the tetrameric <html><a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a></html> construct was commercially </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>synthesized by IDT (Integrated DNA Technologies) and cloned into a pET-plasmide in order to </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>synthesized by IDT (Integrated DNA Technologies) and cloned into a pET-plasmide in order to </div></td></tr>
</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=385514&oldid=prevRomanM89 at 01:57, 18 October 20142014-10-18T01:57:12Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/4/42/MR_transformed-cells.png" alt="scheme" width="47%" /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/4/42/MR_transformed-cells.png" alt="scheme" width="47%" /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption" style="text-align:justify;width: 94%;"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption" style="text-align:justify;width: 94%;"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <b>Figure 2:<del class="diffchange diffchange-inline"></b> </del>Transformation of epithelial-derived cancer cells overexpressing EpCAM. <del class="diffchange diffchange-inline"> </del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <b>Figure 2: Transformation of epithelial-derived cancer cells overexpressing EpCAM.<ins class="diffchange diffchange-inline"></b></ins></div></td></tr>
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</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=385474&oldid=prevRomanM89 at 01:56, 18 October 20142014-10-18T01:56:44Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/8/8d/Mr_cancersurf_1.png" alt="" /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/8/8d/Mr_cancersurf_1.png" alt="" /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 1: (A) Scheme of the DARPin library design. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 1: (A)<ins class="diffchange diffchange-inline"></b> </ins>Scheme of the DARPin library design. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (B) Tetrameric <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>(B)<ins class="diffchange diffchange-inline"></b> </ins>Tetrameric <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> construct. Four DARPin Ec1 molecules were fused via linker peptides to a Streptavidin protein. At the N terminus the Ec1 DARPin is fused to a polyhistidine-Tag (His-Tag) and the C terminus prevents the linkage to the Streptavidin monomer. Adapted from Stumpp et al. 2008.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> construct. Four DARPin Ec1 molecules were fused via linker peptides to a Streptavidin protein. At the N<ins class="diffchange diffchange-inline">-</ins>terminus </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>the Ec1 DARPin is fused to a polyhistidine-Tag (His-Tag) and the C<ins class="diffchange diffchange-inline">-</ins>terminus prevents the linkage to the Streptavidin monomer. </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>Adapted from Stumpp et al. 2008.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/4/42/MR_transformed-cells.png" alt="scheme" width="47%" /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/4/42/MR_transformed-cells.png" alt="scheme" width="47%" /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption" style="text-align:justify;width: 94%;"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption" style="text-align:justify;width: 94%;"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 2: Transformation of epithelial-derived cancer cells overexpressing EpCAM. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 2:<ins class="diffchange diffchange-inline"></b> </ins>Transformation of epithelial-derived cancer cells overexpressing EpCAM. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 3:</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 3:</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Purification of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Size-exclusion chromatogram and </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> Purification of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>.<ins class="diffchange diffchange-inline"></b> </ins>Size-exclusion chromatogram and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> coomassie stained SDS-PAGE of purified <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Upper SDS-PAGE: </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> coomassie stained SDS-PAGE of purified <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Upper SDS-PAGE: </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Fractions taken from Ni-NTA purification. M: marker, L: lysate, FT: flow through, </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Fractions taken from Ni-NTA purification. M: marker, L: lysate, FT: flow through, </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/5/54/Mr_cancersurf_3.png" alt="" /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/5/54/Mr_cancersurf_3.png" alt="" /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 4: (A) Immunofluorescent staining of Caco-2 and 3T3 cells incubated with </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 4: (A)<ins class="diffchange diffchange-inline"></b> </ins>Immunofluorescent staining of Caco-2 and 3T3 cells incubated with </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> protein. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> protein. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Both cell lines were stained with phalloidin (red) conjugated with rhodamine (F-Actin) and Alexa488 (green) conjugated anti-His-antibody </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Both cell lines were stained with phalloidin (red) conjugated with rhodamine (F-Actin) and Alexa488 (green) conjugated anti-His-antibody </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (<a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>) in presence of 25 &micro;M </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (<a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>) in presence of 25 &micro;M </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. The nucleus was stained with DAPI (blue). Scale bar represents 5 µm. (B) </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. The nucleus was stained with DAPI (blue). Scale bar represents 5 µm. <ins class="diffchange diffchange-inline"><b></ins>(B)<ins class="diffchange diffchange-inline"></b> </ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Immunofluorescent staining of A549 cells incubated with </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Immunofluorescent staining of A549 cells incubated with </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> protein. Cells were stained with rhodamine (red) conjugated phalloidin </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> protein. Cells were stained with rhodamine (red) conjugated phalloidin </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> in presence of 25 µM <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. The nucleus was stained with DAPI (blue). The control </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> in presence of 25 µM <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. The nucleus was stained with DAPI (blue). The control </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> was stained in </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> was stained in </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> absence of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Scale bar represents 5 µm. (C) Binding specificity of </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> absence of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Scale bar represents 5 µm. <ins class="diffchange diffchange-inline"><b></ins>(C)<ins class="diffchange diffchange-inline"></b> </ins>Binding specificity of </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> to epithelial-derived cancer cell lines. Fluorescence of Alexa488-coupled </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> to epithelial-derived cancer cell lines. Fluorescence of Alexa488-coupled </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> anti-His-antibody in A549 and Caco-2 (epithelial) cells was normalized to 3T3 (interstitial) cells. (D) Fluorescence intensities of Caco-2 and 3T3 cells in comparison.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> anti-His-antibody in A549 and Caco-2 (epithelial) cells was normalized to 3T3 (interstitial) cells. <ins class="diffchange diffchange-inline"><b></ins>(D)<ins class="diffchange diffchange-inline"></b> </ins>Fluorescence intensities of Caco-2 and 3T3 cells in comparison.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 5: Cellular response after TLR-5 activation by Flagellin. <del class="diffchange diffchange-inline"> </del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 5: Cellular response after TLR-5 activation by Flagellin.<ins class="diffchange diffchange-inline"></b></ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/0/07/Mr_cancersurf_4.png" alt="" /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/0/07/Mr_cancersurf_4.png" alt="" /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 6: Purification of <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> and binding </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 6: Purification of <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> and binding </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> assays with <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (A) Size-exclusion </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> assays with <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (A)<ins class="diffchange diffchange-inline"></b> </ins>Size-exclusion </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> chromatogram of <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> chromatogram of <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> in complex with its chaperone FliS. Upper SDS-PAGE: Ni-NTA purification. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> in complex with its chaperone FliS. Upper SDS-PAGE: Ni-NTA purification. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> M: marker, L: lysate, FT: flow through, W: wash, E: elution. Lower SDS-PAGE: Samples of peak fractions </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> M: marker, L: lysate, FT: flow through, W: wash, E: elution. Lower SDS-PAGE: Samples of peak fractions </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. (B) Streptavidin- and GST-interaction assay using purified </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. <ins class="diffchange diffchange-inline"><b></ins>(B)<ins class="diffchange diffchange-inline"></b> </ins>Streptavidin- and GST-interaction assay using purified </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> and Hag-Strep/FliS. (1) Strep-beads without protein (contaminant protein at 30 kDa). (2) Strep-beads with Hag-Strep/FliS immobilized. (3) </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> and Hag-Strep/FliS. (1) Strep-beads without protein (contaminant protein at 30 kDa). (2) Strep-beads with Hag-Strep/FliS immobilized. (3) </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep/FliS. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep/FliS. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (C) Interaction of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> and </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>(C)<ins class="diffchange diffchange-inline"></b> </ins>Interaction of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> visualized by competition assays. Soluble fractions: </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> visualized by competition assays. Soluble fractions: </div></td></tr>
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<td colspan="2" class="diff-lineno">Line 222:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/e/e0/Mr_cancersurf_5.png" alt="" /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/e/e0/Mr_cancersurf_5.png" alt="" /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 7: Electron micrographs of uranyl-acetate stained <i>B. subtilis</i> cells. (A) Whole cell of wild type strain </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 7: Electron micrographs of uranyl-acetate stained <i>B. subtilis</i> cells. (A)<ins class="diffchange diffchange-inline"></b> </ins>Whole cell of wild type strain </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> 3610 (B) Whole cell of <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> strain. Scale bar represents 1 μm. (C) </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> 3610 <ins class="diffchange diffchange-inline"><b></ins>(B)<ins class="diffchange diffchange-inline"></b> </ins>Whole cell of <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> strain. Scale bar represents 1 μm. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>Magnification of flagellar </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <b></ins>(C)<ins class="diffchange diffchange-inline"></b> </ins>Magnification of flagellar filaments from 3610 wild type strain. Scale bar represents 100 nm <ins class="diffchange diffchange-inline"><b></ins>(D)<ins class="diffchange diffchange-inline"></b> </ins>Magnification of flagellar filaments </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>filaments from 3610 wild type strain. Scale bar represents 100 nm (D) Magnification of flagellar filaments </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> from Hag-D2-Strepstrain. Scale bar represents 300 nm. <ins class="diffchange diffchange-inline"><b></ins>(E)<ins class="diffchange diffchange-inline"></b> </ins>Swarming assay of both <i>B. subtilis</i> strains. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> from Hag-D2-Strepstrain. Scale bar represents 300 nm. (E) Swarming assay of both <i>B. subtilis</i> strains. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>(F)<ins class="diffchange diffchange-inline"></b> </ins>Swimming assay of both <i>B. subtilis</i> strains.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (F)Swimming assay of both <i>B. subtilis</i> strains.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/6/6f/MR_tumor_Terminator.png" alt="" /></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2014/6/6f/MR_tumor_Terminator.png" alt="" /></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <span class="caption"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Figure 8: Model for possible future applications in lung cancer treatment.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><b></ins>Figure 8: Model for possible future applications in lung cancer treatment.<ins class="diffchange diffchange-inline"></b></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One of the strongest interactions established by biological molecules is the interaction </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One of the strongest interactions established by biological molecules is the interaction </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>between Streptavidin and <del class="diffchange diffchange-inline">biotinn</del>, a cofactor of carboxylases. The property of Streptavidin to </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>between Streptavidin and <ins class="diffchange diffchange-inline">biotin</ins>, a cofactor of carboxylases. The property of Streptavidin to </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>tightly bind substrates in combination with engineered Strep-Tags (peptide-tag) has been </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>tightly bind substrates in combination with engineered Strep-Tags (peptide-tag) has been </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>utilized over the years for several approaches, e.g. biochemical and immunohistochemical </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>utilized over the years for several approaches, e.g. biochemical and immunohistochemical </div></td></tr>
</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=384380&oldid=prevRomanM89 at 01:46, 18 October 20142014-10-18T01:46:45Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 01:46, 18 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. (B) Streptavidin- and GST-interaction assay using purified </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. (B) Streptavidin- and GST-interaction assay using purified </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> and Hag-Strep/FliS. (1) Strep-beads without protein (contaminant protein at 30 kDa). (2) Strep-beads with Hag-Strep immobilized. (3) </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> and Hag-Strep/FliS. (1) Strep-beads without protein (contaminant protein at 30 kDa). (2) Strep-beads with Hag-Strep<ins class="diffchange diffchange-inline">/FliS </ins>immobilized. (3) </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep<ins class="diffchange diffchange-inline">/FliS</ins>. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (C) Interaction of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> and </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (C) Interaction of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> visualized by competition assays. Soluble fractions: </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> visualized by competition assays. Soluble fractions: </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (1) Hag-Strep. (2) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> (1) Hag-Strep<ins class="diffchange diffchange-inline">/FliS</ins>. (2) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a>. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (3) Hag-Strep and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (4) </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> (3) Hag-Strep<ins class="diffchange diffchange-inline">/FliS </ins>and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (4) </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> and </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (5) </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (5) </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Insoluble fractions: (6) Hag-Strep and </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Insoluble fractions: (6) Hag-Strep<ins class="diffchange diffchange-inline">/FliS </ins>and </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5). (7) </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5 <ins class="diffchange diffchange-inline">dilution</ins>). (7) </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> and </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> and </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5).(8) Hag-Strep and </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5 <ins class="diffchange diffchange-inline">dilution</ins>).(8) Hag-Strep<ins class="diffchange diffchange-inline">/FliS </ins>and </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1). </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1 <ins class="diffchange diffchange-inline">dilution</ins>). </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (9) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (9) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1).</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1 <ins class="diffchange diffchange-inline">dilution</ins>).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=383894&oldid=prevRomanM89 at 01:42, 18 October 20142014-10-18T01:42:32Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One of the strongest interactions established by biological molecules is the interaction </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One of the strongest interactions established by biological molecules is the interaction </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">of </del>Streptavidin and <del class="diffchange diffchange-inline">biotin</del>, a cofactor of carboxylases. The property of Streptavidin to </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">between </ins>Streptavidin and <ins class="diffchange diffchange-inline">biotinn</ins>, a cofactor of carboxylases. The property of Streptavidin to </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>tightly bind substrates in combination with engineered Strep-Tags (peptide-tag) has been </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>tightly bind substrates in combination with engineered Strep-Tags (peptide-tag) has been </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>utilized over the years for several approaches, e.g. biochemical and immunohistochemical </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>utilized over the years for several approaches, e.g. biochemical and immunohistochemical </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to selectively stain the outer membrane by biotinylated fluorophores.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to selectively stain the outer membrane by biotinylated fluorophores.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Our <del class="diffchange diffchange-inline">goal </del>was to use the extremely tight interaction of Streptavidin and the engineered </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Our <ins class="diffchange diffchange-inline">aim </ins>was to use the extremely tight interaction of Streptavidin and the engineered Strep-TagII, </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Strep-TagII, which binds to Streptavidin in the low pico molar range, in combination with </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>which binds to Streptavidin in the low pico molar range, in combination with </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>DARPin to selectively target epithelial-derived cancer cells and amplify the interaction by </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>DARPin to selectively target epithelial-derived cancer cells and amplify the interaction by </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>our Strep-tagged flagellar filament scaffold. Therefore, we designed a codon-optimized </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>our Strep-tagged flagellar filament scaffold. Therefore, we designed a codon-optimized </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Streptavidin/DARPin chimeric gene (<html><a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a></html>) </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Streptavidin/DARPin chimeric gene (<html><a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a></html>) </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the heterologous production in ''E. coli''. This chimeric gene enables the efficient </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the heterologous production in ''E. coli''. This chimeric gene enables the efficient </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>production of StrepDARPidin in ''E. coli'' and may also serve <del class="diffchange diffchange-inline">for future iGEM teams </del>as a </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>production of StrepDARPidin in ''E. coli'' and may also serve as a template for the codon optimized Streptavidin gene <ins class="diffchange diffchange-inline">for future iGEM teams</ins>, </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>template for the codon optimized Streptavidin gene, thereby improving the existing </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>thereby improving the existing </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Streptavidin biobrick gene in the registry. Additionally, we established a refolding protocol </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Streptavidin biobrick gene in the registry. Additionally, we established a refolding protocol </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the purification of StrepDARPidin and verified its functionality and selectivity by an </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the purification of StrepDARPidin and verified its functionality and selectivity by an </div></td></tr>
</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=382639&oldid=prevRomanM89 at 01:31, 18 October 20142014-10-18T01:31:02Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></html></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></html></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>CancerSURF combines the scaffold of a tetrameric Streptavidin to increase the specificity of a DARPin to detect a prominent tumor surface marker of many epithelial-derived</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>CancerSURF combines the scaffold of a tetrameric Streptavidin to increase the specificity of a DARPin to detect a prominent tumor surface marker of many epithelial-derived </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>cancer cells (like lung and colon cancer cells), called EpCAM (epithelial cell adhesion molecule; Went et al. 2006). EpCAM is expressed by every </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>cancer cells (like lung and colon cancer cells), called EpCAM (epithelial cell adhesion molecule; Went et al. 2006). EpCAM is expressed by every </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>epithelial cell on the basolateral side in a specific ratio while being overexpressed on the surface irregularly by transformed tumor cells (Figure 2). We planned </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>epithelial cell on the basolateral side in a specific ratio while being overexpressed on the surface irregularly by transformed tumor cells (Figure 2). We planned </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to fuse an EpCAM-binding DARPin Ec1 (Stefan et al. 2011) with a tetrameric Streptavidin increasing the local concentration of the construct for targeting EpCAM-positive </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to fuse an EpCAM-binding DARPin Ec1 (Stefan et al. 2011) with a tetrameric Streptavidin increasing the local concentration of the construct for targeting EpCAM-positive </div></td></tr>
</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=382494&oldid=prevRomanM89 at 01:29, 18 October 20142014-10-18T01:29:39Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></html></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></html></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>CancerSURF combines the scaffold of a tetrameric Streptavidin <del class="diffchange diffchange-inline">(Strep) </del>to increase the specificity of DARPin <del class="diffchange diffchange-inline">Ec1 for detection of </del>a prominent tumor surface marker of many </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>CancerSURF combines the scaffold of a tetrameric Streptavidin to increase the specificity of <ins class="diffchange diffchange-inline">a </ins>DARPin <ins class="diffchange diffchange-inline">to detect </ins>a prominent tumor surface marker of many epithelial-derived</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>epithelial-derived cancer cells (like lung <del class="diffchange diffchange-inline">& </del>colon cancer cells), called EpCAM (<del class="diffchange diffchange-inline">Epithelial Cell Adhesion Molecule) (</del>Went et al. 2006). EpCAM is expressed by every </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins>cancer cells (like lung <ins class="diffchange diffchange-inline">and </ins>colon cancer cells), called EpCAM (<ins class="diffchange diffchange-inline">epithelial cell adhesion molecule; </ins>Went et al. 2006). EpCAM is expressed by every </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>epithelial cell on the basolateral side in a specific ratio while being overexpressed on the surface irregularly by transformed tumor cells (Figure 2). We planned </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>epithelial cell on the basolateral side in a specific ratio while being overexpressed on the surface irregularly by transformed tumor cells (Figure 2). We planned </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to fuse an EpCAM-binding DARPin Ec1 (Stefan et al. 2011) with a tetrameric Streptavidin increasing the local concentration of the construct for targeting EpCAM-positive </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to fuse an EpCAM-binding DARPin Ec1 (Stefan et al. 2011) with a tetrameric Streptavidin increasing the local concentration of the construct for targeting EpCAM-positive </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Al-Khodor S, Price CT, Kalia A, Abu Kwaik Y (2010). Ankyrin-repeat containing proteins of microbes: a conserved structure with functional diversity.Trends Microbiol. 18, 132–139 10.1016</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Al-Khodor S, Price CT, Kalia A, Abu Kwaik Y (2010). Ankyrin-repeat containing proteins of microbes: a conserved structure with functional diversity.Trends Microbiol. 18, 132–139 10.1016</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Howarth M, TingAY (2008).Imaging proteins in live mammalian cells with biotin ligase and monovalent Streptavidin. Nature Protocols 3, 534 - 545 <del class="diffchange diffchange-inline">(2008).</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Howarth M, TingAY (2008). Imaging proteins in live mammalian cells with biotin ligase and monovalent Streptavidin. Nature Protocols 3, 534 - 545 </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Leigh ND, Guanglin Bian, Xilai Ding, Hong Liu, Semra Aygun-Sunar, Lyudmila G. Burdelya, Andrei V. Gudkov, Xuefang Cao (2014). A Flagellin-Derived Toll-Like Receptor 5 Agonist Stimulates Cytotoxic Lymphocyte-Mediated Tumor Immunity. PLoS ONE 9(1): e85587</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Leigh ND, Guanglin Bian, Xilai Ding, Hong Liu, Semra Aygun-Sunar, Lyudmila G. Burdelya, Andrei V. Gudkov, Xuefang Cao (2014). A Flagellin-Derived Toll-Like Receptor 5 Agonist Stimulates Cytotoxic Lymphocyte-Mediated Tumor Immunity. PLoS ONE 9(1): e85587</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Maaser K, Borlak J (2008). A genome-wide expression analysis identifies a network of EpCAM-induced cell cycle regulators.British Journal of Cancer <del class="diffchange diffchange-inline">(2008) </del>99,1635 – 1643<del class="diffchange diffchange-inline">.</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Maaser K, Borlak J (2008). A genome-wide expression analysis identifies a network of EpCAM-induced cell cycle regulators.British Journal of Cancer 99,1635 – 1643</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Stefan N, Martin-Killias P, Wyss-Stoeckle (2011). DARPins Recognizing the Tumor-Associated Antigen EpCAM Selected by Phage and Ribosome Display and Engineered for Multivalency.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Stefan N, Martin-Killias P, Wyss-Stoeckle (2011). DARPins Recognizing the Tumor-Associated Antigen EpCAM Selected by Phage and Ribosome Display and Engineered for Multivalency.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Antonie van Leeuwenhoek 105:275–288</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Antonie van Leeuwenhoek 105:275–288</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Went P, Vasei M, Bubendorf L (2006).Frequent high-level expression of the</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Went P, Vasei M, Bubendorf L (2006). Frequent high-level expression of the Immunotherapeutic targetEp-CAM in colon, stomach, prostate and lung cancers. British Journal of Cancer 94, 128 – 135 </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Immunotherapeutic targetEp-CAM in colon, stomach, prostate and lung cancers. British Journal of Cancer <del class="diffchange diffchange-inline">(2006) </del>94, 128 – 135</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
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</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=381477&oldid=prevRomanM89 at 01:20, 18 October 20142014-10-18T01:20:42Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. (B) Streptavidin- and GST-interaction assay using purified </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. (B) Streptavidin- and GST-interaction assay using purified </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep/FliS</a> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> and Hag-Strep/<del class="diffchange diffchange-inline">FlyS</del>. (1) Strep-beads without protein (contaminant protein at 30 kDa). (2) Strep-beads with Hag-Strep immobilized. (3) </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> and Hag-Strep/<ins class="diffchange diffchange-inline">FliS</ins>. (1) Strep-beads without protein (contaminant protein at 30 kDa). (2) Strep-beads with Hag-Strep immobilized. (3) </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a>. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a>. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a>. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a>. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (C) Interaction of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> and </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (C) Interaction of <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> and </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> visualized by competition assays. Soluble fractions: </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> visualized by competition assays. Soluble fractions: </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (1) Hag-Strep. (2) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a>. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> (1) Hag-Strep. (2) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a>. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (3) Hag-Strep and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (4) </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (3) Hag-Strep and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (4) </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> and </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a> and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (5) </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. (5) </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Insoluble fractions: (6) Hag-Strep and </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a>. Insoluble fractions: (6) Hag-Strep and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5). (7) </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5). (7) </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> and </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a> and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5).(8) Hag-Strep and </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:5).(8) Hag-Strep and </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1). </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1). </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> (9) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> (9) <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> and <a href="http://parts.igem.org/Part:BBa_K1329000" target="_blank">StrepDARPidin</a> (1:1).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </span></div></td></tr>
</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=381094&oldid=prevRomanM89 at 01:17, 18 October 20142014-10-18T01:17:16Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> in complex with its chaperone FliS. Upper SDS-PAGE: Ni-NTA purification. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> in complex with its chaperone FliS. Upper SDS-PAGE: Ni-NTA purification. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> M: marker, L: lysate, FT: flow through, W: wash, E: elution. Lower SDS-PAGE: Samples of peak fractions </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> M: marker, L: lysate, FT: flow through, W: wash, E: elution. Lower SDS-PAGE: Samples of peak fractions </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. (B)Streptavidin-and GST-interaction assay using purified </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> taken from size-exclusion fractions. (B) Streptavidin- and GST-interaction assay using purified </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep<ins class="diffchange diffchange-inline">/FliS</ins></a> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> and Hag-Strep. (1) Strep-beads without protein. (2) Strep-beads with Hag-Strep immobilized. (3) </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> and Hag-Strep<ins class="diffchange diffchange-inline">/FlyS</ins>. (1) Strep-beads without protein <ins class="diffchange diffchange-inline">(contaminant protein at 30 kDa)</ins>. (2) Strep-beads with Hag-Strep immobilized. (3) </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a>. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> Strep-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a>. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a>. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (4) GST-beads with Hag-Strep. (5) GST-beads with <a href="http://parts.igem.org/Part:BBa_K1329003" target="_blank">Hag-D2-Strep</a>. </div></td></tr>
</table>RomanM89http://2014.igem.org/wiki/index.php?title=Team:Marburg:Project:Tumor&diff=379660&oldid=prevRomanM89 at 01:05, 18 October 20142014-10-18T01:05:34Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One of the strongest interactions established by biological molecules is the interaction </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>One of the strongest interactions established by biological molecules is the interaction </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>of <del class="diffchange diffchange-inline">streptavidin </del>and biotin, a cofactor of carboxylases. The property of <del class="diffchange diffchange-inline">streptavidin </del>to </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>of <ins class="diffchange diffchange-inline">Streptavidin </ins>and biotin, a cofactor of carboxylases. The property of <ins class="diffchange diffchange-inline">Streptavidin </ins>to </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>tightly bind substrates in combination with engineered Strep-Tags (peptide-tag) has been </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>tightly bind substrates in combination with engineered Strep-Tags (peptide-tag) has been </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>utilized over the years for several approaches, e.g. biochemical and immunohistochemical </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>utilized over the years for several approaches, e.g. biochemical and immunohistochemical </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>methods for protein purification and immunofluorescent stainings.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>methods for protein purification and immunofluorescent stainings.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>In 2006, the iGEM team from Harvard used <del class="diffchange diffchange-inline">streptavidin </del>to <del class="diffchange diffchange-inline">target </del>''E. coli'' cells. By placing </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>In 2006, the iGEM team from Harvard used <ins class="diffchange diffchange-inline">Streptavidin </ins>to <ins class="diffchange diffchange-inline">stain the outer membrane of </ins>''E. coli'' cells. By placing </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>the <html><a href="http://parts.igem.org/Part:BBa_J36848" target="_blank"><del class="diffchange diffchange-inline">streptavidin</del></a></html> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>the <html><a href="http://parts.igem.org/Part:BBa_J36848" target="_blank"><ins class="diffchange diffchange-inline">Streptavidin</ins></a></html> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>coding sequence downstream of a membrane targeting signal sequence and outer membrane protein </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>coding sequence downstream of a membrane targeting signal sequence and outer membrane protein </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>under control of an IPTG inducible promoter, the heterologous production in ''E. coli'' was achieved </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>under control of an IPTG inducible promoter, the heterologous production in ''E. coli'' was achieved </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to selectively stain the outer membrane by biotinylated fluorophores.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>to selectively stain the outer membrane by biotinylated fluorophores.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Our goal was to use the extremely tight interaction of <del class="diffchange diffchange-inline">streptavidin </del>and the engineered </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Our goal was to use the extremely tight interaction of <ins class="diffchange diffchange-inline">Streptavidin </ins>and the engineered </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Strep-TagII, which binds to <del class="diffchange diffchange-inline">streptavidin </del>in the low pico molar range, in combination with </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Strep-TagII, which binds to <ins class="diffchange diffchange-inline">Streptavidin </ins>in the low pico molar range, in combination with </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>DARPin to selectively target epithelial-derived cancer cells and amplify the interaction by </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>DARPin to selectively target epithelial-derived cancer cells and amplify the interaction by </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>our Strep-tagged flagellar filament scaffold. Therefore, we designed a codon-optimized </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>our Strep-tagged flagellar filament scaffold. Therefore, we designed a codon-optimized </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the heterologous production in ''E. coli''. This chimeric gene enables the efficient </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the heterologous production in ''E. coli''. This chimeric gene enables the efficient </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>production of StrepDARPidin in ''E. coli'' and may also serve for future iGEM teams as a </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>production of StrepDARPidin in ''E. coli'' and may also serve for future iGEM teams as a </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>template for the codon optimized <del class="diffchange diffchange-inline">streptavidin </del>gene, thereby improving the existing </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>template for the codon optimized <ins class="diffchange diffchange-inline">Streptavidin </ins>gene, thereby improving the existing </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">streptavidin </del>biobrick gene in the registry. Additionally, we established a refolding protocol </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Streptavidin </ins>biobrick gene in the registry. Additionally, we established a refolding protocol </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the purification of StrepDARPidin and verified its functionality and selectivity by an </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>for the purification of StrepDARPidin and verified its functionality and selectivity by an </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>immunofluorescent analysis of targeted cancer cells and Flagellin-strep binding assays, </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>immunofluorescent analysis of targeted cancer cells and Flagellin-strep binding assays, </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Al-Khodor S, Price CT, Kalia A, Abu Kwaik Y (2010). Ankyrin-repeat containing proteins of microbes: a conserved structure with functional diversity.Trends Microbiol. 18, 132–139 10.1016</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Al-Khodor S, Price CT, Kalia A, Abu Kwaik Y (2010). Ankyrin-repeat containing proteins of microbes: a conserved structure with functional diversity.Trends Microbiol. 18, 132–139 10.1016</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
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