[Expression and identification of recombinant Staphylococcus aureus enterotoxin A]

Area of Science:

• Molecular Biology
• Immunology
• Microbiology

Background:

• Staphylococcus aureus enterotoxin A (SEA) is a major cause of foodborne illness.
• Accurate detection and control of SEA are crucial for public health.

Purpose of the Study:

• To clone the SEA gene and construct an expression vector for recombinant SEA (rSEA).
• To achieve high-level expression and purification of rSEA.
• To evaluate the immunogenicity and reactogenicity of the produced rSEA.

Main Methods:

• Synthesized the SEA gene and inserted it into the pET28a prokaryotic expression vector.
• Transfected the recombinant vector into E. coli BL21 (DE3) and induced protein expression using IPTG.
• Purified rSEA using Ni-NTA His Bind affinity chromatography.
• Analyzed protein purity and molecular mass using SDS-PAGE.
• Confirmed specificity using Western blot and evaluated immunogenicity in mice via ELISA.

Main Results:

• Successfully obtained and purified rSEA with a molecular mass of approximately 30 kDa.
• Western blot confirmed specific binding to anti-SEA antibodies.
• ELISA demonstrated that rSEA is immunogenic and reactogenic in BALB/c mice.

Conclusions:

• The constructed rSEA expression vector enabled successful high-level expression of the rSEA protein.
• The purified rSEA exhibits immunogenicity and reactogenicity, suitable for antibody production and immunological detection studies.