Coding

Part:BBa_K4164007

Designed by: Bo Cheng   Group: iGEM22_NAU-CHINA   (2022-09-30)


Ssr

Ssr tag is a peptide sequence from Escherichia coli. It targets the modified protein for degradation, assuring protein-quality control by preventing the accumulation of aberrant, unfinished proteins.

ClpXP and ClpAP degrade ssrA-tagged substrates regardless of their inherent thermodynamic stabilities, whereas FtsH only degrades unstable or metastable ssr-tagged substrates. For example, when a ssr tag is placed at the C-terminus of green-fluorescent protein, which is very stable to denaturation, the resulting protein (GFP-ssr) is degraded efficiently by ClpXP and ClpAP in vitro but is not degraded by FtsH.

In our project, we employed tobacco etch virus protease (TEV) and protein degradation tag Ssr to integrate a logic operation to control the degradation of targeted proteins (mRFP) and resolve the leakage problem. The C-terminus of targeted protein was linked with a protease cleavage site (TEVENLYFQ-protease site) and a degradation tag Ssr (AANDENYAAV) in turn by linkers. When TEV was inductively expressed, it could recognize and cleave specifically at the cleavage site, and the Ssr-tagged proteins that lost the degradation tag were no longer degraded and began to work.

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]


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Categories
Parameters
None