Difference between revisions of "Part:BBa K1722004"
 
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__NOTOC__
 
__NOTOC__
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<html>
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<img style="width:10%" src="https://static.igem.org/mediawiki/2015/2/2c/SZU_China_igem_logo.png"/
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</html>
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{| style="color:black" cellpadding="5" cellspacing="1" border="3" align="right"
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! colspan="2" style="background:#d7474e;"|aminoacyl-tRNA
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|-
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|'''Function'''
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|tRNA
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|-
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|'''Use in'''
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|E.coli&Mammalian cells
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_10 RFC 10]
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|-
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|'''Backbone'''
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|[https://parts.igem.org/Help:Plasmid_Backbones pSB1C3]
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|-
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|'''Submitted by'''
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|[http://2015.igem.org/Team:SZU_China SZU_China 2015]
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|}
 
<partinfo>BBa_K1722004 short</partinfo>
 
<partinfo>BBa_K1722004 short</partinfo>
  
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This is a kind of aminoacyl-tRNA, which has a diffeent structure compared with classic tRNA.The tRNA can only carry a specific amino acid, and is non natural amino acids.It has special anticodons(CUA),which can identify the termination codon(UAG), and then adds the unnatural amino acid carried own into the polypeptide chain to.So that the position of the termination codon corresponds to the insertion of an unnatural amino acid, which allows the protein to continue to be translated.
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The tRNA has a different structure compared with classical tRNA.It 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.
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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.
 
In our project, the aminoacyl-tRNA  was selected by directed evolution.
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===The structure of tRNA===
 
===The structure of tRNA===
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Aminoacyl-tRNA only carries a specific amino acid---- Acetyllysine(ACK),and compared with the classical tRNA has an unusual secondary structure.The structure shows in Fig.1.adds in the protein by Semantic coding of termination codon(UAG)
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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)
 
[[File:TRNA.png|300px|center]]
 
[[File:TRNA.png|300px|center]]
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===Acetyllysine(ACK) & Lysine===
 
===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).
 
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).
 
[[File:Ack.png|500px|center]]
 
[[File:Ack.png|500px|center]]
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
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<partinfo>BBa_K1722004 parameters</partinfo>
 
<partinfo>BBa_K1722004 parameters</partinfo>
 
<!-- -->
 
<!-- -->
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===Design Notes===
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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.
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===Source===
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The original source of this tRNA gene is from barchaebacteria.
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We get this part from Shenzhen Second People's Hospital.
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===References===
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[1]Ibba M．Aminoacyl-tRNA synthetase in archaca：different but not unique．Molecular Microbiology，2003，48(3)：631—637
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[2]Ibba M，Soll D．Aminoacyl-tRNA synthesis．Anna Rev Biochem，2000，69：617--650

Latest revision as of 14:17, 17 September 2015

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 has a different structure compared with classical tRNA.It 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