Fig. 1

Constructs used in this study. a Modified FMDV 2A peptide sequence used in this study. Red arrow represents the site of ribosomal skipping. The long blue arrow represents the peptide that gets added to the C-terminal end of the upstream protein. The short blue arrow represents the amino acid (proline) that gets added to the N-terminal end of the downstream protein. b Bicistronic gene constructs for testing expression of Cel7A and eGFP protein in T. reesei. Cel7A and eGFP are separated by FMDV 2A peptide sequence in either orientation (left panel and right panel). Upon transcription of the bicistronic gene construct, a single mRNA transcript containing eGFP and Cel7A are produced, which upon translation gets cleaved after the glycine residue within the 2A peptide to yield two independent proteins. 21 amino acids of the 2A peptide get added to the protein upstream of the peptide and one amino acid (proline) gets added to the downstream protein. Horizontal arrows and the numbers 1, 2, and 3, below the gene cassettes, represent PCR products that were used to confirm the presence of individual genes in the transformants. c Plasmid maps of the two constructs used in this study. The restriction site, SbfI, represents the site of linearization of the plasmid. Eno enolase, PGK1 phosphoglycerate kinase, HPT hygromycin phosphotransferase, term terminator