title: [Preparation and identification of immune serum against recombinant fusion protein of rhoptry 2 and major surface protein 1 from Toxoplasma gondii].
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology of Wenzhou Medical College, Wenzhou 325032, China.
authors.forename: Wen-shu
authors.initials: WS
authors.lastname: Li
authors.forename: Shan-li
authors.initials: SL
authors.lastname: Zhu
authors.forename: Peng-fei
authors.initials: PF
authors.lastname: Wang
authors.forename: Li-fang
authors.initials: LF
authors.lastname: Zhang
authors.forename: Tai-shan
authors.initials: TS
authors.lastname: Min
authors.forename: Wei-da
authors.initials: WD
authors.lastname: Huang
mesh_terms: 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*
chemical_list: D000913:Antibodies, Protozoan; D007106:Immune Sera; D008565:Membrane Proteins; D015800:Protozoan Proteins; C090555:ROP 2 protein, Toxoplasma gondii; D011993:Recombinant Fusion Proteins