Fig. 4: Shuffling library design and assembly for Spl proteases.
From: Evolution of protease activation and specificity via alpha-2-macroglobulin-mediated covalent capture
a Flow cytometry of Spl protease-displaying S. cerevisiae cells probed with substrates A2M-VWLY ↓ S, -WELQ ↓ S and -RWLL ↓ T (representative data of n = 2 independent inductions and flow cytometry measurements). b SCHEMA algorithm output for Spl proteases SplA-F run multiple times, varying parameters number of cross-overs or minimum block size. c Structural alignment of SplA (PDB: 2W7S), SplB (PDB: 2VID) and SplD (PDB: 4INK) with SCHEMA blocks highlighted by colour. d Cloning strategy of SCHEMA library: Each Spl gene was ordered with intervening dual type IIs restriction sites for Golden Gate assembly. The shuffled Spl genes were cloned into the yeast display vector pYD2. FITC Fluorescein isothiocyanate, Strep-PE Streptavidin-Phycoerythrin. Source data is provided as a Source Data file for e.
