http://2013.igem.org/wiki/index.php?title=Team:Wisconsin-Madison/parts_3&feed=atom&action=historyTeam:Wisconsin-Madison/parts 3 - Revision history2024-03-29T10:00:04ZRevision history for this page on the wikiMediaWiki 1.16.5http://2013.igem.org/wiki/index.php?title=Team:Wisconsin-Madison/parts_3&diff=155753&oldid=prevCbjohnson4 at 21:47, 23 September 20132013-09-23T21:47:20Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 21:47, 23 September 2013</td>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p class = "classtheoverview"> <strong><del class="diffchange diffchange-inline">Taq Ligase</del></strong></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p class = "classtheoverview"> <strong><ins class="diffchange diffchange-inline">Text Plasmids</ins></strong></p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p align="left" class = "classtheinlinecontent2"><del class="diffchange diffchange-inline">Taq Ligase </del>is a <del class="diffchange diffchange-inline">thermostable DNA ligase, which joins adjacent 5' and 3' ends of DNA, removing nicks </del>in <del class="diffchange diffchange-inline">double stranded DNA, active from 45-65 degrees Celsius</del>(<del class="diffchange diffchange-inline">Barany, F. (1991). Proc. Natl. Acad. Sci. USA. 88, 189-193</del>). The <del class="diffchange diffchange-inline">Taq ligase gene is within a kanamycin resistant PET-ALM1 plasmid, downstream of the T7 promoter, upstream of the T7 terminator. Thus, it </del>can be used with <del class="diffchange diffchange-inline">an inducible T7 expression system to express </del>the <del class="diffchange diffchange-inline">enzyme. The protein has an N-Terminus hexa-histidine tag</del>, <del class="diffchange diffchange-inline">joined to </del>the <del class="diffchange diffchange-inline">wild type structure by a thrombin cleavage motif</del>. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p align="left" class = "classtheinlinecontent2"><ins class="diffchange diffchange-inline">In addition to the Gibson enzyme plasmids, we have also provided test plasmids that can be used to test a gibson reaction. The product test plasmid </ins>is a <ins class="diffchange diffchange-inline">UV green fluorescent protein from a p70RG plasmid inserted </ins>in <ins class="diffchange diffchange-inline">a pMSBmCherry </ins>(<ins class="diffchange diffchange-inline">RFP</ins>) <ins class="diffchange diffchange-inline">plasmid backbone</ins>. The <ins class="diffchange diffchange-inline">two parts </ins>can be <ins class="diffchange diffchange-inline">amplified by PCR from their respective plasmids and then </ins>used <ins class="diffchange diffchange-inline">in a gibson assembly reaction. Colonies </ins>with the <ins class="diffchange diffchange-inline">PCR products will be UV Green fluorescent when induced with IPTG if the assembly is correct and false positives will have much lower UV fluorescence</ins>, the <ins class="diffchange diffchange-inline">p70RG false positives having no color and the pMSBmCherry false positives being noticeably red</ins>.</p></div></td></tr>
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</table>Cbjohnson4http://2013.igem.org/wiki/index.php?title=Team:Wisconsin-Madison/parts_3&diff=129757&oldid=prevCbjohnson4: Created page with "<!-- *** What falls between these lines is the Alert Box! You can remove it from your pages once you have read and understood the alert *** --> {{Template:Wisconsin-Madison}} <h..."2013-09-16T23:54:17Z<p>Created page with "<!-- *** What falls between these lines is the Alert Box! You can remove it from your pages once you have read and understood the alert *** --> {{Template:Wisconsin-Madison}} <h..."</p>
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<p class = "classtheoverview"> <strong>Taq Ligase</strong></p><br />
<p align="left" class = "classtheinlinecontent2">Taq Ligase is a thermostable DNA ligase, which joins adjacent 5' and 3' ends of DNA, removing nicks in double stranded DNA, active from 45-65 degrees Celsius(Barany, F. (1991). Proc. Natl. Acad. Sci. USA. 88, 189-193). The Taq ligase gene is within a kanamycin resistant PET-ALM1 plasmid, downstream of the T7 promoter, upstream of the T7 terminator. Thus, it can be used with an inducible T7 expression system to express the enzyme. The protein has an N-Terminus hexa-histidine tag, joined to the wild type structure by a thrombin cleavage motif. </p><br />
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