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> | ||
| − | This | + | This part encodes for the aminoacyl tRNA synthetase (aaRS) for the non-natural amino-acid <i>O</i>‑methyl‑<span style="font-variant:small-caps">l</span>‑tyrosine.<br> |
<br> | <br> | ||
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 a template. | 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 a 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"> | |
| − | + | <p align="left" style="width: 650px; margin-left: 15px; margin-right: 15px;" alt=""> | |
| − | + | <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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| − | </div> | + | </div> |
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| − | The | + | The aaRS is used in combination with the supressor tRNA (for example <a href="https://parts.igem.org/Part:BBa_K1976023">BBa_K1976023). 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>‑methyl‑<span style="font-variant:small-caps">l</span>‑tyrosine. |
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<td style="padding: 0cm 5.4pt; vertical-align: top; width: 306.7pt; height: 214.9pt;"> | <td style="padding: 0cm 5.4pt; vertical-align: top; width: 306.7pt; height: 214.9pt;"> | ||
| − | SDS-PAGE | + | 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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[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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Revision as of 13:18, 22 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.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 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.
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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
[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