Fig. 1: Library design, synthesis and experimental outline.
From: Experimental characterization of de novo proteins and their unevolved random-sequence counterparts
a, Schematic illustration of the in silico design of libraries of de novo and unevolved random-sequence proteins. A de novo library (DN) was built from putative de novo proteins identified in human and fly. Subsequently, a library of unevolved random sequences (R) was designed to mirror the length and amino acid frequencies of library DN. The two libraries were synthesized by oligonucleotide library synthesis ready for experimental study. b, Approaches used to profile solubility and structure content of each library. Following amplification, each library was expressed in a chaperone-assisted cell-free format and structural content was quantified using a proteolytic assay. In parallel, subcloned libraries were expressed in E. coli to screen for soluble and folded variants that did not disrupt periplasmic export. Created with BioRender.com.
