Bioinformatics Pilot Program Abstract: Mark Blenner

Use of High-Throughput Sequencing to Understand Protein Evolution
Dr. Mark Blenner, Department of Chemical and Biomolecular Engineering, Clemson University

The broad, long-term objective of the proposed research is to develop a detailed understanding of how selective pressure influences the evolution of protein-ligand interactions. This knowledge could then be applied in the design of proteins with improved or novel properties. Specifically, we will study the evolution of a modelbacterial transcription regulator, pBAD, as it evolves a new binding specificity while losing the original. We hypothesize that the duration of positive and negative selections is a critical factor for the evolutionary trajectory of protein binding selectivity. We further hypothesize that a controlled evolutionary pressure will in fact result in a repeatable evolutionary trajectory. We will test these hypothesesthrough the following specific aims:Specific Aim 1: Engineer AraC to regulate transcription in response to salicylic acid andnot in response to L-arabinose using several independent continuous directed evolutionexperiments.Specific Aim 2: Use different combinations of positive and negative selection duration toengineer AraC to regulate transcription in response to SA and not in response to arausing continuous directed evolution.