title: Bioinformatics analysis and expression of a novel protein ROP48 in Toxoplasma gondii.
abstract: Toxoplasma gondii is an obligate intracellular apicomplexan parasite, and can infect warmblooded animals and humans all over the world. In the past years, ROP family genes encoding particular proteins of T. gondii had made a great contribution to toxoplasmosis. In this study, we used multiple bioinformatics approaches to predict the physical and chemical characteristics, transmembrane domain, epitope, and topological structure of the rhoptry protein 48 (ROP48). The results indicated that ROP48 protein was mainly located in the membrane and had several positive linear-B cell epitopes and Th-cell epitopes, which suggested that ROP48 is a potential DNA vaccine candidate against toxoplasmosis. Then the PCR product amplified from the ROP48 cDNA was inserted into a pEASY-T1 vector to build a recombinant cloning plasmid. After sequencing, ROP48 was subcloned into a eukaryotic expression plasmid pEGFP-C1 to obtain pEGFP-C1-ROP48 (pROP48). After identification by PCR and restriction enzyme digestion, the recombinant plasmid pROP48 was transfected into HEK 293-T cell and identified by RT-PCR. The results showed that the eukaryotic expression plasmid pROP48 was constructed and transfected to the cells of HEK 293-T successfully. Western blotting showed that the expressed proteins can be recognized by anti-STAg mouse sera.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhou
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.forename: Aihua
authors.initials: A
authors.lastname: Zhou
authors.forename: Gang
authors.initials: G
authors.lastname: Lu
authors.forename: Qihang
authors.initials: Q
authors.lastname: Li
authors.forename: Zhilin
authors.initials: Z
authors.lastname: Wang
authors.forename: Meiyan
authors.initials: M
authors.lastname: Zhu
authors.forename: Huaiyu
authors.initials: H
authors.lastname: Zhou
authors.forename: Hua
authors.initials: H
authors.lastname: Cong
authors.forename: Shenyi
authors.initials: S
authors.lastname: He
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology*; D002460:Cell Line; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods*; D018985:Epitopes, B-Lymphocyte / Q000235:genetics; D018984:Epitopes, T-Lymphocyte / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D008807:Mice, Inbred BALB C; D010957:Plasmids; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology*; D016052:Protozoan Vaccines / Q000008:administration & dosage / Q000276:immunology; D014122:Toxoplasma / Q000235:genetics*; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D008565:Membrane Proteins; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D019444:Vaccines, DNA