Team:Aachen/Project/FRET Reporter
From 2014.igem.org
(→The REACh Construct) |
|||
Line 12: | Line 12: | ||
<li> | <li> | ||
- | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/ | + | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#fluorescence" style="color:black"> |
<div class="menusmall-item menusmall-info" ></div> | <div class="menusmall-item menusmall-info" ></div> | ||
<div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | ||
Line 21: | Line 21: | ||
<li> | <li> | ||
- | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter" style="color:black"> | + | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#fret" style="color:black"> |
<div class="menusmall-item menusmall-info" ></div> | <div class="menusmall-item menusmall-info" ></div> | ||
<div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | ||
Line 30: | Line 30: | ||
<li> | <li> | ||
- | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/ | + | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#darkquencher" style="color:black"> |
<div class="menusmall-item menusmall-info" ></div> | <div class="menusmall-item menusmall-info" ></div> | ||
<div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | ||
Line 39: | Line 39: | ||
<li> | <li> | ||
- | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/ | + | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#gfp-reach" style="color:black"> |
<div class="menusmall-item menusmall-info" ></div> | <div class="menusmall-item menusmall-info" ></div> | ||
<div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | ||
Line 48: | Line 48: | ||
<li> | <li> | ||
- | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter" style="color:black"> | + | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Project/FRET_Reporter#tevprotease" style="color:black"> |
<div class="menusmall-item menusmall-info" ></div> | <div class="menusmall-item menusmall-info" ></div> | ||
<div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | <div class="menusmall-item menusmall-img" style="background: #102030; norepeat scroll 0% 0% transparent; background-size:100%"> | ||
Line 116: | Line 116: | ||
==Cutting the Fusion Protein with the TEV Protease== | ==Cutting the Fusion Protein with the TEV Protease== | ||
- | <span class="anchor" id=" | + | <span class="anchor" id="tevprotease"></span> |
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. | 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. |
Revision as of 07:13, 13 October 2014
|
|
|
|
|