Rotation 1: Mapping the evolutionary trajectories of newly evolved minimal proteins

BBSRC strategic theme: Understanding the rules of life

After millions of years of evolution, small, promiscuous polypeptides have developed into their current forms - large proteins finely tuned towards their function. However, this large form factor makes the study of their evolution difficult, due to the plethora of epistatic effects. Using high-throughput microfluidics-powered directed evolution, this project will study the evolvability of a small mini-cAMPase, previously evolved from a random library using this method. Here, I will study whether the evolutionary trajectories of such a minimal protein differ from more complex, highly evolved and larger proteins observed today. Towards this end, I will record the 'fitness landscapes' of this protein - large datasets of sequence and functional data obtained through the use of microfluidics and next generation sequencing after each round of directed evolution. Ultimately, this will offer us insight into whether the precursors of current large proteins underwent differing evolutionary strategies compared to the evolution observed in their current versions.