FURI | Summer 2020

Phage Assisted Continuous Evolution (PACE) with PhiX174

Phage assisted continuous evolution (PACE) is the most current and advanced method of protein engineering, allowing for accelerated development and complex functions. The research team sought to explore opportunities for PACE with a new phage, PhiX174 and new mechanisms of positive and negative selection. For positive selection, Gene H was chosen because previous literature shows phage viability depends on relative expression levels. A negative selector of MbcT/MbcA was chosen, which counters NAD+ phosphorylation, an essential cofactor for metabolism. The combination of these selectors and our phage will allow for researchers an open access to methods of directed evolution.