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Revision as of 10:05, 7 September 2015

This is a kind of aminoacyl-tRNA(aa-tRNA).

SV40
Function Promoter
Use in Mammalian cells
RFC standard RFC 10
Backbone pSB1C3
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]

This is a kind of aminoacyl-tRNA, which has a different structure compared with classic 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)

TRNA.png

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).

Ack.png

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]


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