Figure 1.

Construction of the recombinant plasmids used for DNA – (a) and protein-delivery (b). Both approaches use the same plasmid backbone consisting of an E. coli origin of replication (oriE1), repD and repE as genes needed for listerial replication, a erythromycin resistance cassette (ermAM) and the gene needed for the balanced lethal system trpS with its own promoter (P_trpS). (a) The genes egfp, deoD, and fcu1 were inserted under the control of the P_CMV or P_4XTETP promoters and fused to a poly-adenylation signal. The autolysis cassette consists of the phage lysin gene ply118 under the control of the listerial promoter P_actA. (b) The E. coli-derived gene deoD is put under the control of P_actA and genetically fused to the signal sequence (SP) for Sec-dependent secretion of ActA.