Part:BBa_K2916009
EF-Tu protein equipped with a 6x HIS affinity tag
Elongation Factor Tu 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 Tu in particular mediates the entry of the aminoacyl-tRNA into a free site of the ribosome
In our part, besides the sequence encoding for the protein we also have a hexahistidine-tag to allow us purify the protein.
In our project we used EF-Tu 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. In fact EF-Tu accounts for 47% of the final protein concentration on the OnePot PURE protein solution.
Characterization
All the data regarding the characterization of that part can be found in the composite part: BBa_K2916038 .
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1183
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 280
Illegal AgeI site found at 685 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 430
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