Difference between revisions of "Part:BBa K2684004"
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Using sfGFP–spycatcher protein(<a href="https://parts.igem.org/Part:BBa_K2684004">Part: BBa_K2684004</a>), 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).  
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Using sfGFP–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).  
 
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<p><img src="https://static.igem.org/mediawiki/2018/5/57/T--SHSBNU_China--21002.jpg" style="width:50%"/></image></p>
 
<p><img src="https://static.igem.org/mediawiki/2018/5/57/T--SHSBNU_China--21002.jpg" style="width:50%"/></image></p>

Revision as of 12:08, 17 October 2018


SpyCatcher-sfGFP

SpyCatcher fused with sfGFP by 2xGGGGS linker

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 514
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 568


Reaction stock leftover in experiment

Using sfGFP–spycatcher protein, the combing function of Spytag and spycatcher system on the biofilm was tested. Gene csgA on the plasmid of pET28a was transferred in to ΔMG1655 as control group. Gene csgA – Spytag on the plasmid of pET28a was transferred in to ΔMG1655 as experiment. To verify the function of Spytag on csgA, the experiment was design to compare the combing rate of sf-GFP – spycatcher protein with cells that have csgA – SpyTag (Experiment) or csgA (Control).

Link: Protocol for SpyTag-SpyCatcher system verification

As can be seen from the result,

Thus we can confirm our csgA – SpyTag system is functional.