Fig. 2

a The ATMT plasmid pTS57 was designed for efficient insertion of genes of interest and screening through Golden Gate Cloning upon replacement with a GFP-drop-out cassette. The gene of interest is expressed with the native T. aurantiacus gpd promotor and xlnR terminator, the hph gene is expressed with the native T. aurantiacus tef-1 promotor and trpC terminator. b PCR analysis to verify the hph integration into T. aurantiacus via ATMT. Optimization of the ATMT procedure for c membrane used, and d incubation time and pH. e A combination of optimized pH and temperature was tested regarding transformation rates. (c: 1 biological replicate, d: 2 biological replicates, and e: 3 biological replicates). Error bars indicate the standard deviation of 3 biological replicates