Part:BBa_K2916008
EF-G protein equipped with a 6x HIS affinity tag
Elongation Factor G is a protein that is used in the PURE and OnePot PURE cell-free systems.
Usage and Biology
Elongation factors are a set of proteins that function during the second step of protein biosynthesis, the elongation phase. They act upon the ribosome, to facilitate translational elongation from formation of the first to the last peptide bond of a growing polypeptide. The elongation phase is the most rapid step in translation, so the factors also regulate the procedure happening during that step to ensure high accuracy of the translation.
The Elongation Factor G in particular catalyzes the translocation of the tRNA and mRNA down the ribosome at the end of each round of polypeptide elongation.
In our part, besides the sequence encoding for the protein we also have a hexahistidine-tag to allow us purify the protein.
For our project we used EF-G 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_K2916039 .
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2113
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 100
Illegal AgeI site found at 538
Illegal AgeI site found at 1882 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 691
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