Difference between revisions of "Part:BBa K2684006"

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<h2>Period - <i>CsgA - SpyTag</i></h2>
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<h2>Overview</h2>
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<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/b/b9/T--SHSBNU_China--21000.png" style="width:75%"/></image></p>
 
<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>  
 
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>
 
 
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Revision as of 19:36, 17 October 2018

CsgA-SpyTag

CsgA fused with SpyTag by 2xGGGGS linker

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Overview

The CsgA sequence was improved from Part BBa_K1583000. We added a SpyTag sequence which fused after csgA gene, creating csgA-spytag (BBa_K2684006). With SpyTag, CotA laccase can be fixed onto the biofilm by forming a covalent bond SpyTag-SpyCatcher.

Gene csgA found in the genome of MG1655 wild type is capable of forming biofilm. Using CRISPR, we knocked out gene csgA on MG1655’s genome creating ΔMG1655 strain. The cell ΔMG1655 would then be used as chassis cell. Gene csgA was fused into plasmid pET28a.