Fig. 1

The expression cassette for C-terminal (a) and N-terminal (b) anchoring of CCL3Gag in L. plantarum. All parts of the cassette are easily exchangeable using restriction sites: SalI between the signal peptide or the N-terminal anchor and CCL3Gag, EcoRI, at the end of the insert, and MluI between CCL3Gag and the C-terminal anchor. a C-terminal anchoring was accomplished by fusing the N-terminus of the CCL3Gag fragment to a signal peptide (SP; from Lp_0373) and the C-terminus to a cell wall anchor (Cwa) from Lp_2578 (Cwa comprises 194 residues of Lp_2578) [42]. b Four N-terminal anchoring motifs were used, all containing an SP. Two lipoanchors were generated using lipobox fragments from Lp_1261 (residues 1–75) and Lp_1452 (residues 1–142), one transmembrane anchor was generated by fusing CCL3Gag to C-terminally truncated Lp_1568 (complete protein with 7-residues truncation), which contains an SP without a predicted signal peptide cleavage site, and one LysM anchor was generated by fusing CCL3Gag to full length Lp_3014 (a 204 residue putative transglycosylase with an N-terminal LysM domain)