A host-plasmid balancing system composed with a delta asd mutant (FaD) of an avirulent strain (T32) of Shigella flexneri 2a and plasmid harboring asd gene was used to express enterotoxigenic E. coli surface antigen 6(CS6) and V. cholerae toxin subunit B (CTB). The results of Western blotting and ELISA showed that all of recombinant plasmids (pYX201, pYX202 and pYX203) could be maintained stably and expressed CS6 and CTB respectively in T32 without any antibiotic selection. All the recombinant bacterial strains could elicit the corresponding antibodies in rabbits. The antibodies against CTB elicited by both FaD/pYX201 and FaD/pYX203 showed to be high level, and had long prolongation time, in otherwise, the antibodies against CS6 showed to be low level, indicating that higher expression level of foreign antigen may be benefit for construction of genetic multivalent vaccine.