A shuttle plasmid, pKSET-CrfP, was constructed to induce the production of BGs in Streptococcus suis (SSBGs) upon anhydrotetracycline induction.The screened WR12, a cell-penetrating peptide (CCP), significantly enhanced the production of SSBGs and achieved a 100% inactivation for vaccine preparation. The SSBGs vaccine exhibited excellent protective efficacy against S. suis challenge in a mouse model.