Difference between revisions of "Part:BBa K2684000"
Line 21: | Line 21: | ||
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). | 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). | ||
</p> | </p> | ||
− | <p><img src="https://static.igem.org/mediawiki/2018/5/57/T--SHSBNU_China--21002.jpg" | + | <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 | Link: Protocol for <a href="http://2018.igem.org/Team:SHSBNU_China/Protocal#SSS">SpyTag-SpyCatcher</a> system verification | ||
</p> | </p> |
Revision as of 11:02, 17 October 2018
CotA Laccase of B.subtilis
CotA Laccase is an endospore type protein secreted from B.subtilis
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1348
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 286
Period - CsgA - SpyTag
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).
As can be seen from the result,
Thus we can confirm our csgA – SpyTag system is functional.
References
Guan, Z.-B., Luo, Q., Wang, H.-R., Chen, Y., & Liao, X.-R. (2018). Bacterial laccases: promising biological green tools for industrial applications. Cellular and Molecular Life Sciences.