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Production and Characterization of MIC1: A Lectin from Toxoplasma gondii

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Lectin Purification and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2132))

Abstract

Some lectins of pathogens interact with host cells through the recognition of specific carbohydrates displayed on the mammals’ cell surface. The microneme protein 1 (MIC1) from Toxoplasma gondii has a lectin domain that specifically binds sialic acid residues, often found in the terminal positions of N-glycans of mammalian cells. The necessary studies on the MIC1 biological roles have been limited initially by the laborious purification of the protein from T. gondii tachyzoites and the low yields verified. Then Escherichia coli has been transformed with a construct containing the MIC1 gene, and the obtained recombinant MIC1 (rMIC1) has been purified from the inclusion bodies. Herein, we detail the methodology of heterologous production and purification of rMIC1 and protocols to assay the rMIC1 lectin ability.

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Authors and Affiliations

  1. Laboratory of Immunochemistry and Glycobiology, Department of Cell and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto Medical School, University of São Paulo (FMRP/USP), Ribeirão Preto, SP, Brazil

    Flávia Costa Mendonça-Natividade, Rafael Ricci-Azevedo & Maria Cristina Roque-Barreira

  2. Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil

    Sandra Maria de Oliveira Thomaz

Authors
  1. Flávia Costa Mendonça-Natividade
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  2. Rafael Ricci-Azevedo
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  3. Sandra Maria de Oliveira Thomaz
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  4. Maria Cristina Roque-Barreira
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Corresponding author

Correspondence to Maria Cristina Roque-Barreira .

Editor information

Editors and Affiliations

  1. Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan

    Jun Hirabayashi

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Costa Mendonça-Natividade, F., Ricci-Azevedo, R., de Oliveira Thomaz, S.M., Roque-Barreira, M.C. (2020). Production and Characterization of MIC1: A Lectin from Toxoplasma gondii. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_38

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  • DOI: https://doi.org/10.1007/978-1-0716-0430-4_38

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0429-8

  • Online ISBN: 978-1-0716-0430-4

  • eBook Packages: Springer Protocols

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