Protein_Domain
TEV clv.

Part:BBa_K1362453

Designed by: Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha   Group: iGEM14_Heidelberg   (2014-10-08)
Revision as of 09:19, 21 October 2019 by Studentwang (Talk | contribs)


TEV protease recognition/cleavage site

This part codes for the amino acid sequence ENLYFQG. This site is recognized by TEV protease (catalytic domain of the Tobacco Etch Virus nuclear inclusion a (NIa) protein), which will cleave the Gln-Gly peptide bond [1]. The final four nucleotides of this sequence are GGGT, which will be the overhang produced wenn a reverse-facing BsaI site (BBa_K1362423, BBa_K1362427, BBa_K1362447) is directly following this part, as in the Sortase A circularization constructs (BBa_K1362202, BBa_K1362203, BBa_K1362204, BBa_K1362205).

  • TEV Protease is a recombinant protease derived from Tobacco Etch Virus (TEV) Nla, which is used to excise the affinity tag of the purified fusion protein. TEV protease has strong site specificity and can recognize the seven amino acid sequence of EXXYXQ(G/S) (Glu-Asn-Leu- Tyr-Phe-Gln-Gly). The most common one is ENLYFQG, and its cleavage site is at Between glutamine and glycine or serine.

Experimental Results

T--LZU-CHINA--TEVp.png
  • Using the specificity of TEVp, we cleave the recognition sites of ALKBH (Fig.A) and Casp3 (Fig.B) linked by the target sequence, which are active.

Usage and Biology

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