Part:BBa_K2916003
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 2989
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1737
Illegal AgeI site found at 471
Illegal AgeI site found at 2859 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 958
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