Part:BBa_K1976020

O-Methyl-L-tyrosine tRNA synthetase

This Part encodes the tRNA for non-natural amino-acid O-Methyl-L-Tyrosine and its synthetase.

The orthogonal pair from the "Expanded Genetic Code Measurement Kit" by the iGEM team Austin Texas 2014 was used as a template.

Figure 1:Dimer of the Methanocaldococcus jannaschii tyrosyl-tRNA synthetase specific for O-methyl-l-tyrosine

Usage

The orthogonal pair for amber supression consists of the tRNA with an anticodon complementary to the amber stop codon and an aminoacyl RNA synthetase (BBa_K1976021) specifically loading the tRNA with the non-natural amino acid. The occurence of O-methyl-L-tyrosine does not stop the translation of the immunoprotein.

Characteristica

SDS-PAGE of E. coli TOP10 culture lysate after 6 hours of constitutive expression of OMT-RS. Left: Cell lysate from E. coli TOP10 not transformed with any plasmid. Right: Cell lysate from E. coli TOP10 transformed with the constitutive OMT generator BBa_K1976022. The OMT-RS holds a molar mass of ~35 kDa.

Figure 2: SDS Page result

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]

References
[1] L. Wang, J. Xie and P. G. Schultz, Expanding the genetic code, Annu Rev Biophys, vol. 35, pp. 225-249, 2006
[2] L. Wang, A. Brock, B. Herberich and P. G. Schultz, Expanding the genetic code of Escherichia coli, Science, vol. 292, pp.498-500, 2001
[3] C. C. Liu and P. G. Schultz, Adding new chemistries to the genetic code, Annu Rev Biochem, vol. 79, pp.413-444, 2010
[4] Y. Zhang, L. Wang, P. G. Schultz and I. A. Wilson, Crystal structures of apo wild-type M. jannaschii tyrosyl-tRNA synthetase (TyrRS) and an engineered TyrRS specific for O-methyl-L-tyrosine, Protein Sci, vol. 14, pp.1340-1349, 2005