Bioinformatics Analysis, Cloning And Expression Of A Novel Toxoplasma Gondii Protein TgIMP1

Toxoplasma gondii is an obligate intracellular parasitic protozoa that can infect a wide variety of mammals and humans. T.gondii infected adults are mostly latent infection, without showing obvious symptoms; while immunodeficiency, immunosuppression or organ transplant patients infected with T.gondii can cause a variety of diseases and may be fatal. T.gondii infection in pregnant women can be transmitted vertically through the placenta to the fetus, causing birth defects, stillbirth, or result in the survival of fetal malformations or mental stunting. T.gondii infection of livestock will also make a serious economic loss.As a new kind of vaccine, the synthesis of DNA vaccine is simple, easy to operate, and better immunity attracted great attention. In recent years, the study of DNA vaccine against T. gondii has made a great progress; however, few vaccines have completely controlled toxoplasmosis. This study analyzed and identied a novel T. gondii protein termed immune mapped protein1(TgIMP1). This kind of protective antigen protein was first discovered in Eimeria spp by Blake et al., and Blake et al., also believes that’homologues of the IMP1antigen, which is protective against Eimeria maxima infection, can be identified in T. gondii and Neospora caninum; which suggest that there may be some characteristic(s) of protective antigens shared across this diverse group of parasites’. Therefore, we are confident to infer the TgIMP1also has the ability of anti-T. gondii infection. We used multiple bioinformatics approaches to predict the physical and chemical characters, signal peptide, transmembrane domain, epitope, topological structure and function of the protein. The result suggested that TgIMP1gene ORF length of1,697bp, encoding a protein containing400amino acids, and the protein containing a plurality of post-translational modification sites, hydrophilic regions, the transmembrane sequence and lymphoid B cell antigen epitope, which theoretically defined that TgIMP1potentially immunogenic, and suggesting that the TgIMP1may be a vaccine candidate against toxoplasmosis. Then we cloned and expressed the TgIMP1gene in HFF cells, the results showed that the eukaryotic expression plasmid pBudCE4.1-TgIMP1was constructed and transfected to the HFF cells successfully.Our study not only proves TgIMP1antigen and immune protection, not only from the technical to prove as the feasibility of anti-toxoplasmosis DNA vaccine, also introduced a new method-bioinformatics analysis method in the study of Toxoplasma pave the way for later Toxoplasma gondii.