First round of directed
evolution of A. flos-aquae and B. megaterium structural proteins.
(A, B) Diagrams of the mutagenized A. flos-aquae and B. megaterium gene clusters,
depicting the scanning site saturation libraries screened in the first
round of evolution. (C, D) Nonlinear US difference signal-to-background
ratio (SBR) from all screened mutants of both clusters. Red lines
indicate the nonlinear difference SBR of the WT for that cluster.
Error bars represent standard error. N = 3 technical
replicates of one biological sample. (E, F) Nonlinear US difference
SBR for the WT and top mutants for each cluster. Error bars represent
standard error. N = 4 biological samples (each an
average of 3 technical replicates). (G, H) Locations of top mutations
from (E, F) in the GvpA/GvpB structure (PDB 8GBS and 7R1C).