Team:Edinburgh/Bacterial/Blue light sensor

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<center><p><br><img src="http://2009.igem.org/wiki/images/a/a3/Strickland.jpg" width="640" height="505" border="0"/></p>
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<center><p><br><img src="http://2010.igem.org/wiki/images/5/59/Ed10-Strickland.jpg" width="640" height="505" border="0"/></p>
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<p>Taken from the article of Strickland et al. (2)</p><br></center>
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<p>Taken from the article of Strickland and al. (2)</p><br></center>
<p>The figure above shows the whole process regarding how the light sensor works, from the dark state (A) to the lgith-activated state (B → C) and then returning to the stable state (D → A). In the dark state, the shared helix contacts the LOV2 domain, and the inactivated TrpR dissociates from the DNA; in the light state, the LOV2 domians absorb the blue light proton and form a covalent adduct between the FMN cofactor and a conserved cysteine residue, destroying the shared helix in the LOV domain and binding / populating an active formation of the TrpR domain. This in turn leads to LovTPR binding the DNA and repressing λcI. However, this binding is not stable, and thus it will eventually return to the initial state.</p>
<p>The figure above shows the whole process regarding how the light sensor works, from the dark state (A) to the lgith-activated state (B → C) and then returning to the stable state (D → A). In the dark state, the shared helix contacts the LOV2 domain, and the inactivated TrpR dissociates from the DNA; in the light state, the LOV2 domians absorb the blue light proton and form a covalent adduct between the FMN cofactor and a conserved cysteine residue, destroying the shared helix in the LOV domain and binding / populating an active formation of the TrpR domain. This in turn leads to LovTPR binding the DNA and repressing λcI. However, this binding is not stable, and thus it will eventually return to the initial state.</p>
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<p>Problems: None yet&nbsp;:)</p>
<p>Problems: None yet&nbsp;:)</p>
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<p>BioBricks: BBa_K191006 (LovTAP), BBa_K191009 (Mutated LovTAP (ILE427 -&gt; PHE)), BBa_K191007 (Trp promoter), etc.
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<p>BioBricks: BBa_K191006 (LovTAP), BBa_K191009 (Mutated LovTAP (ILE427 -&gt; PHE)), BBa_K191007 (Trp promoter), etc.</p>
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Strickland, D., K. Moffat, et al. (2008). "Light-activated DNA binding in a designed allosteric protein." Proceedings of the National Academy of Sciences 105(31): 10709-10714.</p>
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Revision as of 10:28, 6 August 2010







The blue-light sensor, which is composed of the LovTAP hybrid protein designed by Strickland and made available by Prof. Sosnick, was biobricked by EPF-Lausanne in iGEM 2009. It is based on a α-helical domain linker between the LOV2 domain (the photoactive protein) and the E-coli trp repressor, which acts as a conduit for allosteric signals. The effective response of the sensor is at a wavelength of 470nm (as documented by the aforementioned Lausanne iGEM team.

The blue-light sensor (LovTAP) consists of five parts:

  1. Photoreceptor1 (the shared helix between LOV domian and TrpR domain)
    • Dark blue when contacting the LOV domain (dark state).
    • Red when contacting the TrpR domain (light state).
  2. Photoreceptor2 (falvin monoucleotide-FMN cofactor)
  3. LOV domain-orange (photoactive protein)
  4. TrpR domain-grey (DNA regulator)
  5. Operator DNA


Taken from the article of Strickland and al. (2)


The figure above shows the whole process regarding how the light sensor works, from the dark state (A) to the lgith-activated state (B → C) and then returning to the stable state (D → A). In the dark state, the shared helix contacts the LOV2 domain, and the inactivated TrpR dissociates from the DNA; in the light state, the LOV2 domians absorb the blue light proton and form a covalent adduct between the FMN cofactor and a conserved cysteine residue, destroying the shared helix in the LOV domain and binding / populating an active formation of the TrpR domain. This in turn leads to LovTPR binding the DNA and repressing λcI. However, this binding is not stable, and thus it will eventually return to the initial state.

Problems: None yet :)

BioBricks: BBa_K191006 (LovTAP), BBa_K191009 (Mutated LovTAP (ILE427 -> PHE)), BBa_K191007 (Trp promoter), etc.