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Licensed Unlicensed Requires Authentication Published by De Gruyter April 13, 2018

Structural predication and antigenic analysis of Toxoplasma gondii ROP20

Jian Zhou, Wanchun Wang, Pengxia Song, Lin Wang, Yali Han, Jingjing Guo, Zhen Hao, Xi Zhu, Qiong Zhou, Xiadong Du, Gang Lu, Shenyi He and Yingquan Luo
From the journal Acta Parasitologica


Toxoplasma gondii infects almost all the warm-blooded animals. ROP20 protein is expressed in the rhoptry of Toxoplasma gondii. In this study, the secondary structure of ROP20 was analyzed using SMART software. We constructed and analyzed the 3D model of ROP20 protein using SWISS-MODEL online procedure and Visual Molecular Dynamics (VMD) software. The structure analysis fully indicated that ROP20 protein is an important member of the ROP family. Furthermore, We used DNASTAR software and Epitope Database online service to analyze liner-B cell epitopes and T-cell epitopes of ROP20 protein. All the analysis results of ROP20 protein can provide positive information on treatment and vaccine for toxoplasmosis. Moreover, ROP20 gene was obtained from PCR, and a recombinant eukaryotic expression vector (pEGFP-C1-ROP20) was constructed in the following study. After restriction enzyme digestion, the constructed plasmid was transfected into HEK 293-T cells. The RT-PCR result indicated that the recombinant plasmid could transcribe successfully in HEK 293-T cell. The results of western blotting indicated the expressed proteins can be recognized by anti-STAg mouse sera.


This work was supported by the Fundamental Research Funds for the Central Universities of Central South University, the Central South University Sports Medicine Scholarship, the National Natural Science Foundation of China (Grant Nos. 81071373 and81271857), the Shandong Provincial Natural Science Foundation (Grant No. ZR2016HM74) and the National College Students’ Innovation and Entrepreneurship Training Program (Grant No. 201710422116).

  1. Conflict of interest

    Conflict of interests: The authors wish to declare that there is no known conflict of interests associated with this publication.


Bai Y., He S., Zhao G., Chen L., Shi N. Zhou H. et al. 2012. Toxoplasma gondii: bioinformatics analysis, cloning and expression of a novel protein TgIMPl. Experimental Parasitology, 132, 458–464. 10.1016/j.exppara.2012.09.015Search in Google Scholar PubMed

Bradley P.J., Ward C., Cheng S.J., Alexander D.L., Coller S. Coombs G. H. et al. 2005. Proteomic analysis of rhoptry organelles reveals many novel constituents for host-parasite interactions in Toxoplasma gondii. The Journal of Biological Chemistry, 280, 34245–3425810.1074/jbc.M504158200Search in Google Scholar PubMed

Cao A., Liu Y., Wang J., Li X., Wang S. Zhao Q. et al. 2015. Toxoplasma gondii: Vaccination with a DNA vaccine encoding Tand B-cell epitopes of SAG1, GRA2, GRA7 and ROP16 elicits protection against acute toxoplasmosis in mice. Vaccine,33, 6757–6762. 10.1016/j.vaccine.2015.10.077Search in Google Scholar

Constant S., Pfeiffer C., Woodard A., Pasqualini T. Bottomly K. 1995. Extent of T cell receptor ligation can determine the functional differentiation of naive CD4+ T cells. The Journal of Experimental Medicine, 182, 1591–159610.1084/jem.182.5.1591Search in Google Scholar PubMed

Cui Y., Yi L., Zhang M.M., Zhou Y., Tan Z.G. Huang T. et al. 2017. WITHDRAWN: Knockdown of LncRNA HOXA11 Antisense Promotes Glioma Cell Apoptosis Via Sponging MiR-140-5p. Oncology Research, [Epub ahead of print]. 10.3727/096504017X14972669062547Search in Google Scholar

Han Y., Zhou A., Lu G., Zhao G., Wang L. Guo J. et al. 2017. Protection via a ROM4 DNA vaccine and peptide against Toxoplasma gondii in BALB/c mice. BMC Infectious Diseases, 17, 59. 10.1186/s12879-016-2104-zSearch in Google Scholar

Hopp T.P. Woods K.R. 1981. Prediction of protein antigenic determinants from amino acid sequences. Proceedings of the National Academy of Sciences of the United States of America,78, 3824–382810.1073/pnas.78.6.3824Search in Google Scholar PubMed

Liu K.Y., Zhang D.B., Wei Q.K., Li J., Li G.P. Yu J.Z. 2006. Biological role of surface Toxoplasma gondii antigen in development of vaccine. World journal of gastroenterology, 12, 2363–236810.3748/wjg.v12.i15.2363Search in Google Scholar PubMed

Lu G., Wang L., Zhou A., Han Y., Guo J. Song P. et al. 2015. Epitope analysis, expression and protection of SAG5A vaccine against Toxoplasma gondii. Acta Tropica, 146, 66–72. 10.1016/j.actatropica.2015.03.013Search in Google Scholar

Lu G., Zhou A., Meng M., Wang L., Han Y. Guo J. et al. 2014. Alpha-galactosylceramide enhances protective immunity induced by DNA vaccine of the SAG5D gene of Toxoplasma gondii. BMC Infectious Diseases,14, 3862. 10.1186/s12879-014-0706-xSearch in Google Scholar PubMed

Lu G., Zhou J., Zhou A., Han Y., Guo J. Song P. et al. 2017. SAG5B and SAG5C combined vaccine protects mice against Toxoplasma gondii infection. Parasitology International, 66, 596–602. 10.1016/j.parint.2017.06.002Search in Google Scholar PubMed

Nabi H., Rashid I., Ahmad N., Durrani A., Akbar H. Islam S. et al.2017. Induction of specific humoral immune response in mice immunized with ROP18 nanospheres from Toxoplasma gondii. Parasitology Research, 116, 359–37010.1007/s00436-016-5298-5Search in Google Scholar PubMed

Peixoto L., Chen F., Harb O.S., Davis P.H., Beiting D.P. Brownback C.S. et al. 2010. Integrative genomic approaches highlight a family of parasite-specific kinases that regulate host responses. Cell Host & Microbe, 8, 208–218. 10.1016/j.chom.2010.07.004Search in Google Scholar PubMed

Ram H., Rao J.R., Tewari A.K., Banerjee P.S. Sharma A.K. 2013. Molecular cloning, sequencing, and biological characterization of GRA4 gene of Toxoplasma gondii. Parasitology Research, 112, 2487–2494. 10.1007/s00436-013-3414-3Search in Google Scholar PubMed

Romano P., Giugno R. Pulvirenti A. 2011. Tools and collaborative environments for bioinformatics research. Briefings in Bioinformatics, 12, 549–561. 10.1093/bib/bbr055Search in Google Scholar PubMed

Wang H.L., Wang Y.J., Pei Y.J., Bai J.Z., Yin L.T. Guo R. et al. 2016. DNA vaccination with a gene encoding Toxoplasma gondii Rhoptry Protein 17 induces partial protective immunity against lethal challenge in mice. Parasite (Paris, France), 23, 4. 10.1051/parasite/2016004Search in Google Scholar PubMed

Wang H.L., Zhang T.E., Yin L.T., Pang M., Guan L. Liu H.L. et al. 2014. Partial protective effect of intranasal immunization with recombinant Toxoplasma gondii rhoptry protein 17 against toxoplasmosis in mice. PloS One, 9, e108377. 10.1371/journal.pone.0108377Search in Google Scholar PubMed

Wang L., Lu G., Zhou A., Han Y., Guo J. Zhou H. et al. 2016. Evaluation of immune responses induced by rhoptry protein 5 and rhoptry protein 7 DNA vaccines against Toxoplasma gondii. Parasite Immunology, 38, 209–217. 10.1111/pim.12306Search in Google Scholar PubMed

Wang X., Lv G., Li J., Wang B., Zhang Q. Lu C. 2017. LncRNA-RP11-296A18.3/miR-138/HIF1A Pathway Regulates the Proliferation ECM Synthesis of Human Nucleus Pulposus Cells (HNPCs). Journal of Cellular Biochemistry, 118, 4862–4871. 10.1002/jcb.26166Search in Google Scholar PubMed

Wang Y., Wang G., Ou J., Yin H. Zhang D. 2014. Analyzing and identifying novel B cell epitopes within Toxoplasma gondii GRA4. Parasites & Vectors, 7, 474. 10.1186/s13071-014-0474-xSearch in Google Scholar

Wang Y., Wang M., Wang G., Pang A., Fu B., Yin H. et al. 2011. Increased survival time in mice vaccinated with a branched lysine multiple antigenic peptide containing B- and T-cell epitopes from T. gondii antigens. Vaccine, 29, 8619–8623. 10.1016/j.vaccine.2011.09.016Search in Google Scholar

Xiong Y., Wang L., Li Y., Chen M., He W. Qi L. 2017. The Long Non-Coding RNA XIST Interacted with MiR-124 to Modulate Bladder Cancer Growth, Invasion and Migration by Targeting Androgen Receptor (AR). Cellular Physiology and Biochemistry, 43, 405–418. 10.1159/000480419Search in Google Scholar

Yang W.B., Zhou D.H., Zou Y., Chen K., Liu Q. Wang J.L. et al.2017. Vaccination with a DNA vaccine encoding Toxoplasma gondii ROP54 induces protective immunity against toxoplasmosis in mice. Acta Tropica, 176, 427–432. 10.1016/j.actatropica.2017.09.007Search in Google Scholar PubMed

Yu S.Y., Dong B., Zhou S.H. Tang L. 2017a. LncRNA MALAT1: A potential regulator of autophagy in myocardial ischemia-reperfusion injury. International Journal of Cardiology, 247, 25. 10.1016/j.ijcard.2017.04.011Search in Google Scholar

Yu S.Y., Dong B., Zhou S.H. Tang L. 2017b. LncRNA UCA1 modulates cardiomyocyte apoptosis by targeting miR-143 in myocardial ischemia-reperfusion injury. International Journal of Cardiology, 247, 31. 10.1016/j.ijcard.2017.05.055Search in Google Scholar

Zhang T.E., Yin L.T., Li R.H., Wang H.L., Meng X.L. Yin G.R. 2015. Protective immunity induced by peptides of AMA1, RON2 and RON4 containing T-and B-cell epitopes via an intranasal route against toxoplasmosis in mice. Parasites &Vectors, 8, 15. 10.1186/s13071-015-0636-5Search in Google Scholar PubMed

Zhao G., Zhou A., Lu G., Meng M., Sun M. Bai Y. et al. 2013. Identification and characterization of Toxoplasma gondii aspartic protease 1 as a novel vaccine candidate against toxoplasmosis. Parasites & Vectors, 6, 175. 10.1186/1756-3305-6-175Search in Google Scholar PubMed

Zhou J. Wang L. 2017. SAG4 DNA and Peptide Vaccination Provides Partial Protection against T. gondii Infection in BALB/c Mice. Frontiers in Microbiology, 8, 1733. 10.3389/fmicb.2017.01733Search in Google Scholar

Zhou J., Lu G. He S. 2016. Analysis of structures and epitopes of a novel secreted protein MYR1 in Toxoplasma gondii. Folia Parasitologica, 63. 10.14411/fp.2016.028Search in Google Scholar PubMed

Zhou J., Lu G., Wang L., Zhou A.H., Han Y.L. Guo J.J. et al. 2017. Structuraland antigenic analysis of a new Rhoptry Pseudokinase Gene (ROP54) in Toxoplasma gondii. Acta Parasitologica, 62, 513–519. 10.1515/ap-2017-0061Search in Google Scholar PubMed

Zhou J., Wang L., Lu G., Zhou A., Zhu M. Li Q. et al. 2016. Epitope analysis and protection by a ROP19 DNA vaccine against Toxoplasma gondii. Parasite (Paris, France), 23, 17. 10.1051/parasite/2016017Search in Google Scholar PubMed

Zhou J., Wang L., Zhou A., Lu G., Li Q. Wang Z. et al. 2016. Bioinformatics analysis and expression of a novel protein ROP48 in Toxoplasma gondii. Acta Parasitologica, 61, 319–328. 10.1515/ap-2016-0042Search in Google Scholar PubMed

Zuo Z.K., Gong Y., Chen X.H., Ye F., Yin Z.M. Gong Q.N. et al. 2017. TGFbeta1-Induced LncRNA UCA1 Upregulation Promotes Gastric Cancer Invasion and Migration. DNA Cell Biology, 36, 159–167. 10.1089/dna.2016.3553Search in Google Scholar

Received: 2017-11-27
Revised: 2018-1-12
Accepted: 2018-1-18
Published Online: 2018-4-13
Published in Print: 2018-6-26

© 2018 W. Stefański Institute of Parasitology, PAS