Improve

INTRODUCTION

A core principle of the Biobrick foundation is to bring standardisation in a field that is constantly changing and evolving. Standardising biological parts and their assembly allows for projects to focus less on assembly of genetic constructs and more on design and testing. #### LovTAP is a DNA binding fusion protein between a Light-oxygen-voltage-sensing (LOV2) domain from Avena sativa and tryptophan repressor (TrpR) common in E. coli developed and improved in 2008[1],[2]. The LovTAP fusion protein was initially submitted to the registry by the École Polytechnique Fédérale de Lausanne 2009 iGEM team[3]. The LovTAP fusion protein acts as a photoswitch allowing selective DNA binding through the TrpR domain regulated by the asLOV2 domain. The asLOV2 domain sensitive to photoexitation at 470 nm exists in two conformational; a dark state and light state. Photoexcitation at 470 nm of the LOV domain results in the conformational change of the trpR domain allowing DNA binding, DNA disassociation occurs when the LOV domain returns to the inactive dark state causing the conformation to revert.

IMPROVEMENTS

The original Biobrick [http://parts.igem.org/wiki/index.php?title=Part:BBa_K191006 K191006] is only compatible with a single assembly method (RFC 21), limiting availability on the registry and hence hindering further future work. Redesigning of the original brick and optimising the coding sequence for the iGEM registry would result in extended compatibility, facilitating future work and characterisation.

REFERENCES

[1] Strickland, D. et al. ‘Rationally Improving LOV Domain–based Photoswitches’. Nat. Methods 7, 623–626 (2010).

[2] Strickland, D., Moffat, K. & Sosnick, T. R. ‘Light-Activated DNA Binding in a Designed Allosteric Protein.’ Proc. Natl. Acad. Sci. U. S. A. 105, 10709–14 (2008).

[3] ‘Team:EPF-Lausanne/LOVTAP - 2009.igem.org’. Available at: https://2009.igem.org/Team:EPF-Lausanne/LOVTAP.

Gold Medal Criterion #2

Standard Tracks: Improve the function of an existing BioBrick Part. The original part must NOT be from your 2017 part number range. If you change the original part sequence, you must submit a new part. In addition, both the new and original part pages must reference each other. This working part must be different from the part documented in bronze #4 and silver #1.

Special Tracks: Improve the function of an existing iGEM project (that your current team did not originally create) and display your achievement on your wiki.