[Optimizing expression and antibody preparation of recombinant Streptococcus mutans surface protein]

Area of Science:

• Microbiology
• Molecular Biology
• Immunology

Background:

• Streptococcus mutans is a key pathogen in dental caries.
• The surface protein rPAc plays a role in S. mutans virulence.
• Efficient expression systems are needed for studying bacterial surface proteins.

Purpose of the Study:

• To optimize the soluble expression of recombinant Streptococcus mutans surface protein (rPAc) in Escherichia coli.
• To generate a specific polyclonal antibody against rPAc.

Main Methods:

• Optimized culture conditions for rPAc expression in E. coli.
• Purified rPAc was used to immunize mice for polyclonal antibody production.
• Western blot and ELISA were employed to validate antibody specificity and titer.

Main Results:

• Maximal soluble rPAc expression achieved in Luria-Bertani medium (pH 7.2) at 30°C with 1.0 mmol/L IPTG induction.
• The resulting antiserum showed a high titer of approximately 1:6000 via ELISA.
• Western blot confirmed the specific recognition of rPAc by the antibody.

Conclusions:

• Effective soluble expression of rPAc in E. coli was demonstrated.
• A high-specificity polyclonal antibody against rPAc was successfully prepared.
• These findings support further investigation into DNA prime-protein boost inoculation strategies.