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Team:Bonn
From 2013.igem.org
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Making versatile control of biological machines easily possible for everybody we engineer a system for | Making versatile control of biological machines easily possible for everybody we engineer a system for | ||
| - | light dependend control of protein activity per protein degradation. | + | light dependend control of protein activity per protein degradation.</br> |
For protein degradation we harness the ClpXP protease system of prokaryotes. This system enables to | For protein degradation we harness the ClpXP protease system of prokaryotes. This system enables to | ||
activate degradation of a peptid tag (ssrA) fused protein upon induction with the adaptor protein SspB. | activate degradation of a peptid tag (ssrA) fused protein upon induction with the adaptor protein SspB. | ||
Using a Split version of SspB, protein degradation is activated through heterodimerisation of both SspB | Using a Split version of SspB, protein degradation is activated through heterodimerisation of both SspB | ||
| - | parts. | + | parts.</br> |
As a proof of principle we investigate degradation of the fluorescent reporter protein mCherry. | As a proof of principle we investigate degradation of the fluorescent reporter protein mCherry. | ||
To obtain light inducible degradation we use a light dependend heterodimerisation system. The light | To obtain light inducible degradation we use a light dependend heterodimerisation system. The light | ||
| - | sensitive part of this system is LOV2 (A. Sativa). | + | sensitive part of this system is LOV2 (A. Sativa).</br> |
Modelling of the light dependency of protein degradation will even make simple and accurate control of | Modelling of the light dependency of protein degradation will even make simple and accurate control of | ||
protein activity for any desired level of activity possible. | protein activity for any desired level of activity possible. | ||
Revision as of 09:26, 23 June 2013
Regulating protein activity is important throughout lifescience research. Even though there are several
systems available for regulation of proteins all suffer from certain disadvantages.
Aiming to overcome all of these drawbacks we engineer a novel system for
light dependend regulation of protein degradation resulting in a rapid change of protein activity.
Our system can be not only of great importance for fundamental science to investigate protein function but
also for accurate control of bioreactors and biological machines.
Project
Making versatile control of biological machines easily possible for everybody we engineer a system for
light dependend control of protein activity per protein degradation.
For protein degradation we harness the ClpXP protease system of prokaryotes. This system enables to
activate degradation of a peptid tag (ssrA) fused protein upon induction with the adaptor protein SspB.
Using a Split version of SspB, protein degradation is activated through heterodimerisation of both SspB
parts.
As a proof of principle we investigate degradation of the fluorescent reporter protein mCherry.
To obtain light inducible degradation we use a light dependend heterodimerisation system. The light
sensitive part of this system is LOV2 (A. Sativa).
Modelling of the light dependency of protein degradation will even make simple and accurate control of
protein activity for any desired level of activity possible.
Find out more about
split SspB
light inducible heterodimerisation
our protocols
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Background
To enable protein degradation we decided on one proteases system (ClpXP), which allows specific degradation
of proteins, is highly conserved and well established.
For light dependency we compared several systems to finally find the best suited one. We created a
comprehensive overview of all systems making artificially light control possible so that everyone can
easily choose the optimal system for his approach.
Find out more about
the ClpXP protease System
our Comparison of several light sensitive systems
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Human Practice
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Team
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