Composite

Part:BBa_K3147005

Designed by: Thomas Bessede   Group: iGEM19_Montpellier   (2019-10-13)
Revision as of 19:38, 13 October 2019 by Registry (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


TEV protease with TEV-cleavable maltose binding protein

The Montpellier 2019 team made this construction in order to be able to compare the basal activity of the TEV protease (BBa_ K1319004) against the same protease fused to a nanobody. This construction was used as a control within the framework of the KARMA project. This construction produces a TEV merged with an MBP separated by a TEV cutting site (BBa_ J18918). MBP increases the solubility [1] of the fusion protein preventing the aggregation of the protein of interest, this stabilizes expression, the sequence of the produced MBP does not have a signal peptide which allows the protein to be kept in the cytosol. The TEV cutting site allows to separate MBP from the TEV once the protein is produced, indeed the TEV protease is capable of self-cleavage to remove the MBP [2]. An affinity tag 7 histidine is added at the N-ter of the TEV protease and can enable the separation activity of the TEV to be detected by a Western-Blot.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 379
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None