title: Application and expression of Toxoplasma gondii surface antigen 2 (SAG2) and rhoptry protein 2 (ROP2) from recombinant Escherichia coli strain.
abstract: The gene encoding surface antigen 2 (SAG2) or rhoptry protein 2 (ROP2) of Toxoplasma gondii was cloned into the plasmid pGEX-4T-1 and subsequently expressed in Escherichia coli as a glutathione-s-transferase (GST) fusion protein. The characteristics of purified GST-SAG2 or GST-ROP2 were analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis. The specific IgG of a panel of serum samples provided by the National Institute for the Control of Pharmaceutical and Biological Products were tested with commercial ELISA and the lateral flow immunoassay (LFIA) based on GST-SAG2, GST-ROP2 or GST-SAG2+ROP2. A total of 1096 sera and saliva samples from pregnant women were tested by GST-SAG2+ROP2-LFIA. In total, 20 T. gondii IgM positive sera (1.82%), 81 T. gondii IgG positive sera (7.4%) and 23 T. gondii IgA positive saliva (2.1%) were finally confirmed. The SAG2+ROP2 specific IgG and IFN-γ producing CD8+ T cells were induced in mice immunised with GST-SAG2+ROP2. The results indicate that GST-SAG2+ROP2 protein can be used as an antigen for diagnosing T. gondii infection and provide a strategy for development of subunit vaccines for protection against T. gondii infection.
authors.affiliation: Department of Parasitology and Immunology, School of Medicine, Yangzhou University, Yangzhou 225001, China. yzyanhua@yahoo.com.cn
authors.forename: Hua
authors.initials: H
authors.lastname: Yan
authors.forename: Huishen
authors.initials: H
authors.lastname: Yan
authors.forename: Yong
authors.initials: Y
authors.lastname: Tao
authors.forename: Hongju
authors.initials: H
authors.lastname: Chen
authors.forename: Guocai
authors.initials: G
authors.lastname: Li
authors.forename: Weijuan
authors.initials: W
authors.lastname: Gong
authors.forename: Hongmei
authors.initials: H
authors.lastname: Jiao
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.forename: Mingchun
authors.initials: M
authors.lastname: Ji
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000235:genetics / Q000276:immunology*; D000953:Antigens, Protozoan / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007114:Immunization; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000235:genetics / Q000276:immunology* / Q000517:prevention & control; D015800:Protozoan Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D016052:Protozoan Vaccines / Q000276:immunology*; D014122:Toxoplasma / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity; D014123:Toxoplasmosis / Q000235:genetics / Q000276:immunology* / Q000517:prevention & control
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; C067547:surface antigen P22, Toxoplasma