Fig. 3

Demonstration of efficient cleavage of the FMDV 2A peptide in T. reesei. Clonal isolates of the two FMDV 2A-based constructs were compared for their eGFP fluorescence and for their efficiency in expressing Cel7A protein. a Individual clonal isolates were transferred to liquid media and incubated for 3 days at 30 °C in 24-well culture plates. eGFP fluorescence was visualized under blue light. Each of the rows represents five independent clones from a single Cel7A-expressing transformant. +ve and –ve indicate wells containing Cel7A-expressing and Cel7A-deleted transformants, respectively. b Western blotting of secretome from individual wells using anti-Cel7A antibody. c Western blotting of intracellular protein extract from individual clonal isolates using anti-eGFP antibody. Lanes M marker, 1, 2, 3, 4, 5, correspond to individual well coordinates, C2, D2, A4, and A5, represent original Cel7A-expressing transformants from which clonal isolates were generated, JLT102A (AST1116 expressing native Cel7A under the eno promoter), AST1116 (Cel7A-deleted T. reesei QM6a strain)