[Preparation and identification of immune serum against recombinant fusion protein of rhoptry 2 and major surface protein 1 from Toxoplasma gondii].

OBJECTIVE

To prepare and identify immune serum against the recombinant fusion protein of rhoptry 2 (ROP2) and major surface protein 1(P30) from Toxoplasma gondii.

METHODS

The constructed recombinant plasmid of pET28b/ROP2-P30 was transformed to a bacterium BL21-Codon Plus (DE3)-RIL strain and was expressed under IPTG induction. Cells were lysed by multiple rounds of sonication to obtain supernatant and inclusion body respectively. The inclusion body was washed with 2 mol/L and 4 mol/L urea to remove the nonspecific protein. The washed products dissolved in 8 mol/L urea were received by SDS-PAGE. Two rabbits were immunized with the fusion protein rROP2-P30 and sera from the rabbits were collected. Immune diffusion test, indirect ELISA and Western-blot were used to detect antibody titer and specificity of the immune serum against rROP2-P30.

RESULTS

Immune diffusion test demonstrated that specific immune serum were obtained. Indirect ELISA confirmed that the antibody titer in the serum reached 1:12 800 and the rROP2-P30 was recognized by specific IgG in this serum by Western-blot analysis.

CONCLUSION

Specific immune serum against the recombinant fusion protein rROP2-P30 has been prepared.