Part:BBa_K4164006:Design
TEVp
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 461
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
TEVp belongs to the family of C4 peptidases and is structured as a two-domain antiparallel ß-barrel fold, typical of trypsin-like proteases. The TEVp specific recognition motif (TEVp cleavage Site, TS) is only 7 amino acids-long: EXXYXQ-S/G (where X can be any amino acid) and proteolysis takes place between residues Gln and Ser or Gly.
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 (RFP) 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.
Codon optimization for enhanced Escherichia coli expression.
Source
The gene sequence of this part was obtained from NCBI, and its GenBank Accession number is MN658789.1.