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"ROP2"

Original Article
High Expression of Water-Soluble Recombinant Antigenic Domains of Toxoplasma gondii Secretory Organelles
Zhaoshou Yang, Hye-Jin Ahn, Ho-Woo Nam
Korean J Parasitol 2014;52(4):367-376.
Published online August 29, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.4.367

Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with GRA225-105, GRA339-138, ROP2324-561, and MIC21-284 domains had respectively higher value of IgG avidity. The rGST-GRA225-105 and rGST-GRA339-138 were soluble, while rGST-ROP2324-561 and rGST-MIC21-284 were not. GRA231-71, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The rGST-GRA231-71-ROP2324-561, a chimeric protein, appeared to be soluble. Moreover, rGST-GRA231-71-MIC21-284 was also soluble and had higher IgG avidity comparing to rGST-MIC21-284. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.

Citations

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  • Single Cell Expression Systems for the Production of Recombinant Proteins for Immunodiagnosis and Immunoprophylaxis of Toxoplasmosis
    Karolina Sołowińska, Lucyna Holec-Gąsior
    Microorganisms.2024; 12(8): 1731.     CrossRef
  • A vaccine using Anaplasma marginale subdominant type IV secretion system recombinant proteins was not protective against a virulent challenge
    Macarena Sarli, María B. Novoa, Matilde N. Mazzucco, Marcelo L. Signorini, Ignacio E. Echaide, Susana T. de Echaide, María E. Primo, Paulo Lee Ho
    PLOS ONE.2020; 15(2): e0229301.     CrossRef
  • Development of a novel fusion protein with Anaplasma marginale and A. centrale MSP5 improved performance of Anaplasma antibody detection by cELISA in infected and vaccinated cattle
    María E. Primo, Carolina S. Thompson, Beatriz S. Valentini, Macarena Sarli, María B. Novoa, Atilio J. Mangold, Susana T. de Echaide, Ulrike Gertrud Munderloh
    PLOS ONE.2019; 14(1): e0211149.     CrossRef
  • TheToxoplasma gondiidense granule protein TgGRA3 interacts with host Golgi and dysregulates anterograde transport
    Maika S. Deffieu, Tchilabalo Dilezitoko Alayi, Christian Slomianny, Stanislas Tomavo
    Biology Open.2019;[Epub]     CrossRef
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