For construction of the AfuRbp deletion cassette, a sequential cloning strategy was applied as described before. Briefly, 1.5 kb of 5’ and 1.2 kb of 3’ flanking regions of the gene were amplified separately, using RBP_KO1/RBP_KO2 (containing SphI/NcoI restriction sites) and RBP_KO3/RBP_KO4 (containing EcoRI/SalI restriction sites) primer sets, respectively (Table 1). These fragments were cloned next together in pGEM-Teasy vector. At the final step, the A. fumigatus pyrG gene, as a selection marker, with its own promoter and terminator containing EcoRI site at both ends was cloned into EcoRI site, between the 5’and 3’ flanking regions, resulted in pRBP-KO (Fig. 1A).