Difference between revisions of "Part:BBa K3815015"

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==Usage and Biology==
 
==Usage and Biology==
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This is an engineered derivative of wildtype ssrA tag from <i>Escherichia coli</i>, where three C-terminal amino acids LAA in WT are replaced to LGA. To quantify its efficiency of protein degradation, GFP fluorescence intensity of <i>E.coli</i> overnight culture bearing a plasmid expressing GFP-mutant ssrA tag fusion protein was measured by Qubit, and then compared to that of WT and other mutants. Refer to ''<partinfo>BBa_M0050</Partinfo>'' for the biological function of the tag.
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This is an engineered derivative of wildtype ssrA tag from <i>Escherichia coli</i>, where three C-terminal amino acids LAA in WT are replaced to LGA. Refer to ''<partinfo>BBa_M0050</Partinfo>'' for the biological function of the tag.
 
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===Result===
 
===Result===
 
obtained by mutagenizing the WT tag with mixed primers.  
 
obtained by mutagenizing the WT tag with mixed primers.  
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To quantify its efficiency of protein degradation, GFP fluorescence intensity of <i>E.coli</i> overnight culture bearing a plasmid expressing GFP-mutant ssrA tag fusion protein was measured by Qubit, and then compared to that of WT and other mutants.
  
 
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Revision as of 15:01, 20 October 2021


AANDENYALGA. mutant SsrA degradation tag

Usage and Biology

This is an engineered derivative of wildtype ssrA tag from Escherichia coli, where three C-terminal amino acids LAA in WT are replaced to LGA. Refer to BBa_M0050 for the biological function of the tag.


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]


Result

obtained by mutagenizing the WT tag with mixed primers.


To quantify its efficiency of protein degradation, GFP fluorescence intensity of E.coli overnight culture bearing a plasmid expressing GFP-mutant ssrA tag fusion protein was measured by Qubit, and then compared to that of WT and other mutants.



Improvement from existing parts

This is a part improved from BBa_K1051206. In this part, three C-terminal amino acids LAA were replaced with LGA, resulting in a reduced protein degradation efficiency.