Difference between revisions of "Part:BBa K1976020"
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         <h1><i>O</i>-Methyl-<i>L</i>-tyrosine tRNA synthetase </h1>
 
         <h1><i>O</i>-Methyl-<i>L</i>-tyrosine tRNA synthetase </h1>
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This Part encodes the tRNA for nonnatural aminoacid <i>O</i>-Methyl-<i>L</i>-Tyrosine and its synthetase.<br>
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This part encodes for the aminoacyl tRNA synthetase (aaRS) for the non-natural amino-acid <i>O</i>&#8209;methyl&#8209;<span style="font-variant:small-caps">l</span>&#8209;tyrosine.
 
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The orthogonal pair from the <a href="http://2014.igem.org/Team:Austin_Texas/kit">"Expanded Genetic Code Measurement Kit"</a>  by the iGEM team Austin Texas 2014 was used as template.
 
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<img style="width: 40%; height: 40%; margin-left: 15px; margin-right: 15px;" alt="" src="https://static.igem.org/mediawiki/2016/1/17/T--TU_Darmstadt--aaRS.png">
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            <b>Figure 1:</b>Dimer of the <i>Methanocaldococcus jannaschii</i> tyrosyl-tRNA synthetase specific for <i>O</i>-methyl-<span style="font-variant:small-caps">l</span>-tyrosine
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<b>Figure 1:</b>Dimer of the <i>Methanocaldococcus jannaschii</i> tyrosyl-tRNA synthetase specific for <i>O</i>-methyl-<span style="font-variant:small-caps">l</span>-tyrosine.
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The orthogonal pair for amber supression consists of the tRNA with an anticodon complementary to the amber stop codon and an aminoacyl RNA synthetase (<a href="https://parts.igem.org/Part:BBa_K1976021">BBa_K1976021</a>) specifically loading the tRNA with the non-natural amino acid.
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The aaRS is used in combination with the supressor tRNA (for example <a href="https://parts.igem.org/Part:BBa_K1976023">BBa_K1976023</a>). Together they form a so called orthogonal pair used for non-natural amino acid incorporation at position of the <i>amber</i> stop codon. In this case the non-natural amino acid is <i>O</i>&#8209;methyl&#8209;<span style="font-variant:small-caps">l</span>&#8209;tyrosine.
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The occurence of <i>O</i>-methyl-<i>L</i>-tyrosine does not stop the translation of the immunoprotein.
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===<h2>Characteristics</h2>===
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<b>Figure 2:</b> 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 <a href="https://parts.igem.org/Part:BBa_K1976022">BBa_K1976022</a>. The OMT-RS holds a molar mass of ~35 kDa.
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      <img style="width: 300px; height: 500px;" alt="" src="https://static.igem.org/mediawiki/2016/3/35/T--TU_Darmstadt--Synthetase_PAGE.png"></p>
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<h2>Sequence and Features</h2>
 
<h2>Sequence and Features</h2>
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[3] C. C. Liu and P. G. Schultz, Adding new chemistries to the genetic code, Annu Rev Biochem, vol. 79, pp.413-444, 2010<br>
 
[3] C. C. Liu and P. G. Schultz, Adding new chemistries to the genetic code, Annu Rev Biochem, vol. 79, pp.413-444, 2010<br>
 
[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
 
[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
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Latest revision as of 17:38, 23 October 2016

O-Methyl-L-tyrosine tRNA synthetase

This part encodes for the aminoacyl tRNA synthetase (aaRS) for the non-natural amino-acid O‑methyl‑l‑tyrosine.

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


Usage

The aaRS is used in combination with the supressor tRNA (for example BBa_K1976023). Together they form a so called orthogonal pair used for non-natural amino acid incorporation at position of the amber stop codon. In this case the non-natural amino acid is O‑methyl‑l‑tyrosine.

Characteristics

Figure 2: 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.

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