Virulence and antigenicity of the szp-gene deleted Streptococcus equi ssp. zooepidemicus mutant in mice

Streptococcus equi ssp. zooepidemicus (SEZ) causes serious disease in pigs. M-like protein (SzP) of SEZ is a virulence factor with opsonin function. To fabricate better vaccine against SEZ, We have generated a szp-knockout strain by homologous recombination. Two fragments, upstream and downstream of the szp gene, were obtained by genome walking. The vector was then used to delete a 1090 bp segment of szp gene from a strain of SEZ (ATCC35246). The mutant strain had 1000-fold decrease in LD50 as compared with wild type, and provided 85% protection against challenge with a virulent strain when administered as a live vaccine. Semi-quantitative RT-PCR analysis showed a marked increased in levels of IL-4 and IFN-γ mRNA in immunized mice. These data demonstrate that szp-knockout strain is a better candidate for vaccine development.