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Part:BBa_K5150006:Design

Designed by: Lorena Gallegos Solís, Ana Laura Torres Huerta, Alma Paola Castorena Zorrilla, Gabriela Araceli Rosas   Group: iGEM24_TecCEM   (2024-09-30)
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TEV protease recognition sequence for tag removal.


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]


Design Notes

The nucleotide sequence encoding the recognition peptide was generated through reverse translation taking into account as a criterion the codon optimization that was useful for bacteria, both for E. coli and for Bacillus subtillis.


Source

The specific recognition motif for TEV, known as the TEV cleavage site, consists of just 7 amino acids: EXXYXQ-S/G (where X represents any amino acid). Proteolysis occurs between the Gln and Ser or Gly residues. Many studies have demonstrated that within the cleavage site not all residues are equally tolerated. Despite many possible combinations of the cleavage site have been tested, the optimum recognition site is ENLYFQ-S/G, which corresponds to the cleavage site between protein fragments NIb and CP on the natural TEV polyprotein [3]. Despite the sequence of the peptide: ENLYFQS, is identical to the protease's natural substrate peptide, the most commonly use is ENLYFQG [4]. Since a large amount of characterization information has been reported for this sequence, we decided to use it.

References