Figure 2
Construction and characterisation of Listeria-based Mage-b vaccine strains. (A) Three overlapping fragments of mouse Mage-b (homologous to human MAGE-B) were cloned as a fusion protein with a truncated non-cytolytic listeriolysin O (LLO) in the pGG-34 vector under the control of the listerial hemolysin promoter (Phly). (B) Secretion of LLO-Mage-b proteins by the Listeria-based vaccine strains was detected by western blotting using α-myc antibodies (top) and α-pest antibodies (bottom). In the western blot with α-myc antibodies, the LLO-Mage-b proteins are indicated by black arrows, and in the western blot with α-pest antibodies by white arrows. The complete Mage-b protein fused with truncated LLO represents a band of 88 kDa, whereas the three fragments of Mage-b fused with truncated LLO represent a band of 61 kDa. Endogenous LLO (58 KDa) secreted by all LM is indicated by a star.
