Difference between revisions of "Part:BBa K2916008"

Latest revision as of 02:47, 22 October 2019

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2113
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 100
Illegal AgeI site found at 538
Illegal AgeI site found at 1882
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 691

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2916008 short</partinfo>
-Isoleucyl-tRNA synthetase is a fusion protein that is used in the PURE and OnePot PURE cell-free systems.
+Elongation Factor G is a protein that is used in the PURE and OnePot PURE cell-free systems.
+<!-- Add more about the biology of this part here-->
-<!-- Add more about the biology of this part here
 ===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 <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.
+===Characterization===
+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_K2916039"> BBa_K2916039 </a></html>.
 <span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K2916008 SequenceAndFeatures</partinfo>