Synthetic biology approaches to exploring property-function relationships of reflective proteins for adaptive coloration and repellence

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Using modern synthetic biology and recombinant DNA approaches, the aim of this study is to establish methods for (1) the synthesis and characterisation of novel reflectin proteins and (2) the fabrication of reflectin-based films/fibres with desirable optical, structural, and adaptive properties. Achieving these aims will require probing the sequence-property-function relationships of reflectins by firstly rapidly exploring the sequence-structure-functional space using high-throughput assembly, expression and structural/functional characterisation methods, and secondly, designing and utilising methods of fabrication and functional characterisation of reflectin-based films/fibres and photonic structures. Initially, methods for both the hyper-variation of reflectin domains and domain-domain assembly of reflectin sequences into full-length genes will be developed, which will involve predictions of folded structure, estimation/exploration of domain boundaries, and investigations into the role of conserved amino acids in defining optical properties and domain structural integrity. The solubility of reflectin variant domains will then be probed using C-terminal GFP tagging before full-length constructs are formed using automated combinatorial assembly methods and screened for desired solubility. Finally, both the orthogonal introduction of non-canonical amino acids (amino acids possessing novel chemical, physical, and biological properties) will be probed with the aim of introducing novel functionality, likely unattainable by nature, into assembled reflectin domains, followed by the functional analysis of these synthetic reflectin proteins. Methods of fabrication of reflectin-based films and fibres will be explored and developed, for example using spin-, dip, and flow-coating, and sol-gel processing, with the aim of developing materials with desirable properties, particularly adaptive colouration and repellence.