title: Toxoplasma gondii: bioinformatics analysis, cloning and expression of a novel protein TgIMP1.
abstract: Toxoplasma gondii is an obligate intracellular protozoan parasite, infecting a large variety of animals and human beings. In recent years, the study of DNA vaccine against T. gondii has made a great progress; however, few vaccines have completely controlled toxoplasmosis. Thus people started to look for more effective antigenic proteins. Here we report a novel T. gondii protein termed immune mapped protein 1 (TgIMP1). We used multiple bioinformatics approaches to predict the physical and chemical characters, signal peptide, transmembrane domain, epitope, topological structure and function of the protein, and we theoretically determined that the TgIMP1 has multiple epitopes, and with immunogenicity, suggesting that the TgIMP1 may be a vaccine candidate against toxoplasmosis. Then the gene coding TgIMP1 was obtained by PCR and connected with cloning vector. Recombinant plasmid was identified by PCR, double digestion and sequencing analysis. Then the TgIMP1 gene was directly inserted into the eukaryotic expression vector pBudCE4.1, so that the recombinant eukaryotic expression plasmid pBudCE4.1-TgIMP1 was constructed. After identification by PCR and restriction enzyme digestion, the recombinant plasmid pBudCE4.1-TgIMP1 was transfected into cells of HFF, and then identified by RT-PCR. The results showed that the eukaryotic expression plasmid pBudCE4.1-TgIMP1 was constructed and was transfected to the HFF cells successfully.
authors.affiliation: Department of Parasitology, Shandong University School of Medicine, Jinan, Shandong Province 250012, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Bai
authors.forename: Shenyi
authors.initials: S
authors.lastname: He
authors.forename: Guanghui
authors.initials: G
authors.lastname: Zhao
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.forename: Na
authors.initials: N
authors.lastname: Shi
authors.forename: Huaiyu
authors.initials: H
authors.lastname: Zhou
authors.forename: Hua
authors.initials: H
authors.lastname: Cong
authors.forename: Qunli
authors.initials: Q
authors.lastname: Zhao
authors.forename: Xing-Quan
authors.initials: XQ
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D010274:Parasitology / Q000379:methods*; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016052:Protozoan Vaccines / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics / Q000276:immunology*; D019444:Vaccines, DNA / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000953:Antigens, Protozoan; D000939:Epitopes; D021382:Protein Sorting Signals; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D019444:Vaccines, DNA; C577666:immune mapped protein 1, Toxoplasma gondii