Fig. 6

Construction of a PbMAP1-knockout strain (ΔPbMAP1). A Scheme of the transfection plasmid used to target and knock out PbMAP1 in the PbANKA strain using the double-crossover method. B PCR validation of PbMAP1 deletion. PCR products from 5'-UTR, 3′-UTR, and PbMAP1 showed that PbMAP1 was replaced by DHFR. C Western blotting of PbMAP1 expression in WT and ΔPbMAP1. Protein bands with expected molecular weights are shown. No PbMAP1 protein was detected in the knockout strain. GAPDH was used as the control. D Southern blotting using a DNA probe from the PbMAP1 gene in WT and ΔPbMAP1. The gene encoding PbMAP1 was completely deleted in the knockout strain