Fig. 1

Schematic representation of the pJHSP expression vector system. a Schematic representation of fusion proteins JHS02–JHS14 encoded on plasmids pJHS02–pJHS14 that include an N-terminal signal peptide (SP) for SEC-dependent secretion (I.) followed by a StrepII-tag (II.) and the SUMO protein (III.). The protein of interest (POI; IV.) is fused to the C-terminus of SUMO, and to the N-terminus of a 6xHis-tag (V.). c.o.: encoding DNA-sequence codon optimized for B. subtilis. b Map of plasmid pJHS12. Cm: chloramphenicol resistance gene, Amp: ampicillin resistance gene, Rep(pMB1): origin of replication, LacI: Lac repressor gene. The SUMO fusion cassette is controlled by the Pgrac promotor, and a transcriptional terminator (T) sequence is placed at the 3′ end. c Detail of pJHS12 showing relevant sequences of the reporter fusion cassette. A signal peptide-encoding sequence can be inserted in the unique MluI and Eco52I restriction sites for an in frame fusion to the StrepII-SUMO part. A gene of interest can be fused via the unique BamHI, PstI and Eco47III restriction sites with the option to add a C-terminal polyhistidine tag. The SenP-processing site is indicated by scissors. RBS: ribosome binding site