Difference between revisions of "Part:BBa K2684006"
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<partinfo>BBa_K2684006 parameters</partinfo> | <partinfo>BBa_K2684006 parameters</partinfo> | ||
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+ | <html> | ||
+ | <body> | ||
+ | <div> | ||
+ | <h2>Period - <i>CsgA - SpyTag</i></h2> | ||
+ | <p><img src="https://static.igem.org/mediawiki/2018/b/b9/T--SHSBNU_China--21000.png" style="width:75%"/></image></p> | ||
+ | Gene <i>csgA</i> found in the genome of MG1655 wild type is capable of forming biofilm. Using CRISPR, we knocked out gene <i>csgA</i> on MG1655’s genome creating ΔMG1655 strain. The cell ΔMG1655 would then be used as chassis cell. Gene <i>csgA</i> was fused into plasmid pET28a.</p> | ||
+ | The CsgA sequence was improved from <a href="https://parts.igem.org/Part:BBa_K1583000">Part BBa_K1583000</a>. We added a SpyTag sequence which fused after <i>csgA</i> gene, creating <i>csgA-spytag</i> (BBa_K2684006). With SpyTag, CotA laccase can be fixed onto the biofilm by forming a covalent bond SpyTag-SpyCatcher.</p> | ||
+ | <p><img src="https://static.igem.org/mediawiki/2018/8/87/T--SHSBNU_China--21001.jpg" style="width:50%"/></image></p> | ||
+ | <p class="pic_text">Reaction stock leftover in experiment</p> | ||
+ | </div> | ||
+ | <p class="text"> | ||
+ | Using sfGFP(<a href="") – spycatcher protein, the combing function of Spytag and spycatcher system on the biofilm was tested. Gene <i>csgA</i> on the plasmid of pET28a was transferred in to ΔMG1655 as control group. Gene <i>csgA – Spytag</i> on the plasmid of pET28a was transferred in to ΔMG1655 as experiment. To verify the function of Spytag on <i>csgA</i>, the experiment was design to compare the combing rate of sf-GFP – spycatcher protein with cells that have csgA – SpyTag (Experiment) or csgA (Control). | ||
+ | </p> | ||
+ | <p><img src="https://static.igem.org/mediawiki/2018/5/57/T--SHSBNU_China--21002.jpg" style="width:50%"/></image></p> | ||
+ | Link: Protocol for <a href="http://2018.igem.org/Team:SHSBNU_China/Protocal#SSS">SpyTag-SpyCatcher</a> system verification | ||
+ | </p> | ||
+ | <p class="text"> | ||
+ | As can be seen from the result, | ||
+ | </p> | ||
+ | <p class="text"> | ||
+ | Thus we can confirm our <i>csgA – SpyTag</i> system is functional. | ||
+ | </p> | ||
+ | </div> | ||
+ | </body> | ||
+ | </html> |
Revision as of 11:43, 17 October 2018
CsgA-SpyTag
CsgA fused with SpyTag by 2xGGGGS linker
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Period - CsgA - SpyTag
Reaction stock leftover in experiment
As can be seen from the result,
Thus we can confirm our csgA – SpyTag system is functional.