When: December 2, 2015, 2:00 PM
Location: 3rd Floor Orchard View Room , Discovery Building
Contact: 608-316-4401, email@example.com
Design and evolution of new proteins and genomes
The ability to design allosteric TFs to respond to new inducers opens the door to engineering biosynthetic pathways for thousands of valuable molecules that currently lack a natural sensor. However, designing allosteric TFs is a major challenge because it requires engineering a new protein-small molecule interaction without disrupting the allosteric function. Using the classical lac repressor TF (LacI), we developed a general scheme to engineer allosteric TFs responding to new inducers. We used computational protein design to generate candidates, synthesize DNA encoding 20,000 designed variants with chip-based oligonucleotides, and developed a two-step enrichment for allosterically-functional variants inducible by the target ligand. We redesigned LacI to respond to four new inducers: sucralose, lactitol, fucose and gentiobiose, with induction response comparable to the original LacI with IPTG. Besides metabolic engineering applications, this method allows building orthogonal switches for synthetic gene networks, sensors for single cell real-time metabolomics and regulators for targeted control of developmental pathways.
All SysBioM (Systems Biology in Madison) sponsored talks take place on Wednesdays at 2p.m. in the 3rd floor Orchard View room of the Discovery Building. Talks are open to the public. Access to the room is via the elevator behind Aldo’s Cafe in the Northeast corner of the building.