Part:BBa_K4179005

P2A self-cleaving peptide - Optimized for E. coli

• This sequence codes for a P2A, a 2A self-cleaving peptide originating in porcine teschovirus-1. The team of Technion 2022-Israel used the amino acid sequence of P2A studied by Kim et al [1] and found the corresponding DNA sequence that is optimized for E. coli.

• This part was used by the Technion team in six composite systems (BBa_K4179003,Part:BBa_K4179008,Part:BBa_K4179009,Part:BBa_K4179010,Part:BBa_K417913,Part:BBa_K4179014) for the purpose of detection and quantification of the expression of a protein-coding gene. Thus, the P2A sequence was located in-between the gene of interest (GOI) and an mCherry sequence. In such a manner, the mCherry expression is tied directly to the start codon of the GOI while not being covalently bound to it. As P2A has a relatively high self-cleaving efficiency [1], an mCherry signal corresponds to a 1:1 ratio, at the very least, which allows for a lower estimate of the GOI’s expression.

• The GSG amino acids at its N-terminus were added to improve cleavage efficiency, as demonstrated by Kim et al.

• A similar P2A sequence was characterized by the 2014 iGEM team of Warwick (BBa_K1442039).

References

1. Kim JH, Lee SR, Li LH et al. High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. PLoS One 2011; 6.