Exploring the Potential of Networked Directed Evolution Based on Novel LacI/effector Pairs

点击翻译按钮获取中文摘要

Understanding of the evolution of functional proteins remains a daunting challenge, despite intense research efforts in basic and applied research. Nature's ability to create functional proteins is still unrivalled, raising the question in which respect we can can improve biomimetic efforts in directed evolution to make laboratory evolution more powerful. One key difference is the context in which evolution occurs: laboratory evolution usually deals with enzymes as single species, but natural enzymes are typically embedded in regulated networks and interdependent pathways. This project attempts to build simple networks in which expression of a coupled reporter marks E. coli cells that successfully express a new, functional protein and renders them selectable. This approach could expand the reaction classes that are accessible for directed evolutino, as current laboratory evolution is primarily targeted at functions that are assayed directly, e.g. by generating a fluorescent product. We hope to develop a method that will make directed evolution more versatile and efficient, but also provide insight into how network regulation might have contributed to natural evolution.