Difference between revisions of "Part:BBa K2916003"
 
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<partinfo>BBa_K2916003 short</partinfo>
 
<partinfo>BBa_K2916003 short</partinfo>
  
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Glycyl-tRNA synthetase is a fusion protein that is used in the PURE and OnePot PURE cell-free systems.  
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Glycyl-tRNA synthetase is a fusion protein that is used in the OnePot PURE cell-free system.  
  
  
 
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<!-- Add more about the biology of this part here-->
  
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===Usage and Biology===
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===Biology===
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Transfer RNA (tRNA) is the molecule that enables the Genetic Code contained in the nucleotide sequence of a messenger RNA (mRNA) molecule to be translated into the amino acid sequence of a polypeptide chain. This function is catalysed by a group of enzymes called aminoacyl-tRNA synthetase (aaRS) which attach the appropriate amino acid onto its tRNA. This group consists of 20 different types of aminoacyl-tRNA synthetases, one for each amino acid of the genetic code. Those enzymes, in presence of Adenosine triphosphate (ATP) and Amino Acids, produces aminoacyl-tRNA (tRNA charged with an Amino Acid) that can be used by the ribosome to transfer the amino acid from the tRNA to the polypeptide being synthesised, according of course to the genetic code.
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Aminoacyl-tRNA therefore play a major role in RNA translation.
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Glycyl-tRNA synthetase (GlyRS) attaches to Glycine (codons:GGU, GGC, GGA, GGG)
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In our part, besides the sequence encoding for the protein we also have a hexahistidine-tag to allow us purify the protein.
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===Usage===
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In our project we used GlyRS as a part of the protein solution needed for <html><a style="padding: 0px; margin: 0px;" href="https://2019.igem.org/Team:EPFL/OnePot_Pure"> OnePot PURE cell-free system </a></html> using the method of gravity flow affinity chromatography, as described in the <html><a style="padding: 0px; margin: 0px;" href="https://www.protocols.io/view/protein-purification-for-onepot-pure-cell-free-sys-8auhsew"> protocol </a></html> we designed.
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===Characterization===
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All the data regarding the characterization of that part can be found in the composite part: <html><a style="padding: 0px; margin: 0px;" href="https://parts.igem.org/Part:BBa_K2916021"> BBa_K2916021 </a></html>.
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Used in OnePot PURE
 
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2916003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2916003 SequenceAndFeatures</partinfo>

Latest revision as of 01:52, 22 October 2019


GlyRS protein equipped with a 6x HIS affinity tag

Glycyl-tRNA synthetase is a fusion protein that is used in the OnePot PURE cell-free system.


Biology

Transfer RNA (tRNA) is the molecule that enables the Genetic Code contained in the nucleotide sequence of a messenger RNA (mRNA) molecule to be translated into the amino acid sequence of a polypeptide chain. This function is catalysed by a group of enzymes called aminoacyl-tRNA synthetase (aaRS) which attach the appropriate amino acid onto its tRNA. This group consists of 20 different types of aminoacyl-tRNA synthetases, one for each amino acid of the genetic code. Those enzymes, in presence of Adenosine triphosphate (ATP) and Amino Acids, produces aminoacyl-tRNA (tRNA charged with an Amino Acid) that can be used by the ribosome to transfer the amino acid from the tRNA to the polypeptide being synthesised, according of course to the genetic code. Aminoacyl-tRNA therefore play a major role in RNA translation.


Glycyl-tRNA synthetase (GlyRS) attaches to Glycine (codons:GGU, GGC, GGA, GGG)

In our part, besides the sequence encoding for the protein we also have a hexahistidine-tag to allow us purify the protein.


Usage

In our project we used GlyRS as a part of the protein solution needed for OnePot PURE cell-free system using the method of gravity flow affinity chromatography, as described in the protocol we designed.



Characterization

All the data regarding the characterization of that part can be found in the composite part: BBa_K2916021 .



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2989
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1737
    Illegal AgeI site found at 471
    Illegal AgeI site found at 2859
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 958