Related Experiment Video
Updated: May 17, 2026
Toxoplasma gondii Cyst Wall Formation in Activated Bone Marrow-derived Macrophages and Bradyzoite Conditions
Published on: August 12, 2010
[Recombinant expression and immunologic identification of bradyzoite-specific antigen BSR4 of Toxoplasma gondii]
Xing-Zhi Chen1, Kai Guo, Yong Chen
1Department of Microbiology and Parasitology, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 233030, China. chen007835@163.com
Objective:
To express the recombinant BSR4 protein of Toxoplasma gondii Prugniaud (PRU) strain and study its immunologic characteristics.
Methods:
The recombinant plasmid pET28a(+)-BSR4 was transformed into E. coli BL21, followed by expression of BSR4 induced by IPTG and its purification. The immunoreactivity of the recombinant protein BSR4 was analyzed by Western blotting with the sera from mice infected by PRU strain of T. gondii or normal mice as the first antibody. The 3H-TdR incorporation assay was performed to determine the proliferation of splenocytes in mice infected by PRU strain stimulated by BSR4, and the stimulation index (SI) was calculated. ELISA was used to evaluate the immunoreactivity of BSR4 protein, in which 20 sera from each of acute (anti-T. gondii IgG(-)IgM+) and chronic (IgG+IgM(-)) toxoplasmosis patients, and healthy people were tested.
Results:
The recombinant BSR4 was induced and expressed. After denaturation, renaturation and purification, the soluble protein (Mr 45 000) was obtained and detected by anti-T. gondii serum with Western blotting. BSR4 in 1, 5, and 25 microg/ml induced the proliferation of splenocytes in mice infected by PRU strain with higher SI (1.13, 0.88, and 1.17) than that of control (0.46, 0.24, and 0.49, respectively, P<0.01). ELISA showed that the recombinant BSR4 specifically reacted with the sera of toxoplasmosis patients (IgG+IgM(-), 20/20), not with that of the acute cases (IgG(-)IgM+, 0/20).
Conclusion:
The recombinant BSR4 shows specific immunogenicity and immunoreactivity.
