Part:BBa_K1722004
| aminoacyl-tRNA | |
|---|---|
| Function | tRNA |
| Use in | E.coli&Mammalian cells |
| RFC standard | RFC 10 |
| Backbone | pSB1C3 |
| Submitted by | [http://2015.igem.org/Team:SZU_China SZU_China 2015] |
This is a kind of aminoacyl-tRNA(aa-tRNA). The tRNA can only carry a specific amino acid, which is a kind of unnatural amino acids. It has special anticodons(CUA),which can identify the termination codon(UAG), and carry the unnatural amino acid into the polypeptide chain. So that the tRNA that charges the unnatural amino acid can pair with the amber stop codon UAG. The aminoacyl-tRNA came from barchaebacteria, and it is generally applied to unnatural amino acid orthogonal system. In our project, the aminoacyl-tRNA was selected by directed evolution.
The structure of tRNA
Aminoacyl-tRNA carries only a specific amino acid---- Acetyllysine(ACK), which has a different secondary structure with the classical tRNA. The structure shows in Fig.1 adds in the protein by Semantic coding of termination codon(UAG)
Acetyllysine(ACK) & Lysine
Acetyllysine(ACK) is one of unnatural amino acid and only corresponds to a special aminoacyl-tRNA(tRNA). It can be added to the protein by Semantic coding of termination codon(UAG).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The sequence of tRNA gene has only 72 bases, and the traditional PCR method is not applicable, so we get this part by gene synthesising.
Source
The original source of this tRNA gene is from barchaebacteria.
We get this part from Shenzhen Second People's Hospital.
References
[1]Ibba M．Aminoacyl-tRNA synthetase in archaca：different but not unique．Molecular Microbiology，2003，48(3)：631—637
[2]Ibba M，Soll D．Aminoacyl-tRNA synthesis．Anna Rev Biochem，2000，69：617--650
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