Team:Aachen/Project/FRET Reporter
From 2014.igem.org
AZimmermann (Talk | contribs) (→Producing a GFP-REACh Fusion Protein) |
(→Cutting the Fusion Protein with the TEV Protease) |
||
Line 61: | Line 61: | ||
==Cutting the Fusion Protein with the TEV Protease== | ==Cutting the Fusion Protein with the TEV Protease== | ||
- | To cut the GFP_REACh fusion protein | + | To cut the GFP_REACh fusion protein off, we chose '''Tobacco Etch Virus (TEV) protease''', a highly sequence-specific cysteine protease, that is frequently used for the controlled cleavage of fusion proteins ''in vitro'' and ''in vivo''. The native protease also contains an internal self-cleavage site. This site is slowly cleaved to inactivate the enzyme. The physiological reason for the self-cleavage is unknown, however, undesired for our use. Therefore, our team uses a variant of the native TEV protease that contains the mutation S219V which results in an alteration of the cleavage site so that self-inactivation is diminished. |
''Add: [Fig. 5: Picture of protease]'' | ''Add: [Fig. 5: Picture of protease]'' |
Revision as of 15:06, 12 October 2014
|
|
|
|