Fig. 2
Error-prone PCR mutants of LipY-OVA show increased secretion in M. marinum. a Secretion of LipY-OVAwt, LipY-OVA1, LipY-OVA2 and LipY-OVA3 was analyzed by double filter assay. Proteins were visualized by anti-HA. b Sequencing of constructs showed different mutations in both LipY and OVA domains. LipY-OVA1 and LipY-OVA3 show an extension of the HA-tag due to frameshift mutations. c Double filter assay showing secretion of LipY-OVA swap constructs by M. marinum. Mutant LipY and OVA domains were swapped with wild type domains. Total amount of HA-labeled protein was calculated using ImageJ Gel Analysis tools and the highest score was set to 100%