Coding

Part:BBa_K4421010

Designed by: Shuyang Zhu   Group: iGEM22_NMU_China   (2022-06-21)
Revision as of 09:46, 1 July 2022 by Juejue (Talk | contribs)


2A sequence

The self-cleaving 2A peptide sequence, was intercalated between iCASP9 and the GFP tag and it was derived from FMDV (foot-and-mouth disease virus) to allow transcription and expression of one single mRNA molecule.

  • Figure 2: 2A peptide.

Usage and Biology

The 2A self-cleaving peptide was first identified in 1991 from foot-and-mouth disease virus(Ryan et al., 1991). The 2A peptide, containing the canonical motif DxExNPGP (where “x” is any amino acid which is not conserved), induces the recombinant protein containing 2A peptide to shear itself in the cell, often at the site corresponding to the "-NPG/P-" junction(Luke G et al.,2010).In genetic engineering, 2A peptide can cause an open reading frame (ORF) to translate the peptide chain into several independent peptide chains.

advantages

IRES can also translate two peptide chains from an open reading frame,but the peptide chain upstream of IRES is more efficiently expressed than the peptide chain located downstream [10]. In contrast, the peptide chains upstream and downstream of 2A peptide chain exhibit similar amounts in terms of expression efficiency.

reference

Ryan et al., 1991.Cleavage of foot-and-mouth disease virus polyprotein is mediated by residues located within a 19 amino acid sequence.Journal of general virology direct,1991 Nov;72 ( Pt 11):2727-32.https://doi.org/10.1099/0022-1317-72-11-2727

Luke G et al.,2010.2A to the fore - research, technology and applications.Biotechnology & genetic engineering reviews,2010;26:223-60.https://doi.org/10.5661/bger-26-223

Chan HY et al.,2011.Comparison of IRES and F2A-based locus-specific multicistronic expression in stable mouse lines.PloS one,2011;6(12):e28885.https://doi.org/10.1371/journal.pone.0028885


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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Parameters
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