Part:BBa_K3815015
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
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