[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.


CONCLUSIONS
Specific immune serum against the recombinant fusion protein rROP2-P30 has been prepared.
Li
Wen-shu
WS
Department of Microbiology and Immunology of Wenzhou Medical College, Wenzhou 325032, China.
Zhu
Shan-li
SL
Wang
Peng-fei
PF
Zhang
Li-fang
LF
Min
Tai-shan
TS
Huang
Wei-da
WD
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood / Q000276:immunology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera / Q000276:immunology / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D015800:Protozoan Proteins / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D014122:Toxoplasma / Q000235:genetics / Q000276:immunology*
D000913:Antibodies, Protozoan; D007106:Immune Sera; D008565:Membrane Proteins; D015800:Protozoan Proteins; C090555:ROP 2 protein, Toxoplasma gondii; D011993:Recombinant Fusion Proteins