Protein Production

Protein production is induced by the FNR promoter. Each of the 8 proteins listed below can be expressed once a hypoxic environment is sensed. These proteins are all rich in Arginine and Lysine amino acids, which are known to be good CEST candidates.

The proteins that the CEST protein production device can generate are:

• Human Protomine 1 Optimized ([http://parts.igem.org/Part:BBa_K1123013 BBa_K1123013])
• 1ETF ([http://parts.igem.org/Part:BBa_K1123014 BBa_K1123014])
• 1PJN ([http://parts.igem.org/Part:BBa_K1123021 BBa_K1123015])
• 1G70 ([http://parts.igem.org/Part:BBa_K1123016 BBa_K1123016])
• Poly Arginine-Glycine ([http://parts.igem.org/Part:BBa_K1123018 BBa_K1123018])
• Poly Arginine-Serine ([http://parts.igem.org/Part:BBa_K1123019 BBa_K1123019])
• Poly Threonine-Lysine ([http://parts.igem.org/Part:BBa_K1123020 BBa_K1123020])
• Poly Lysine-Serine ([http://parts.igem.org/Part:BBa_K1123021 BBa_K1123021])

These proteins were selected using the RCSB Protein Data Bank. A python program was designed by us alongside a molecular dynamics simulator, to ensure that the selected proteins had a high content of Arginine and Lysine aminoacids. The complete description of the protein selection process can be found on the protein selection section of the drylab tab. The rest of the proteins were selected using the results of McMahon's research (2008) [1].

References

[1] McMahon, M.T., 2008, New “Multi-Color” Polypeptide DIACEST Contrast Agents for MR Imaging, Magn Reson Med. 2008 October ; 60(4): 803–812.