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In vitro generation of cloned populations of Toxoplasma gondii

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Abstract

Cloned populations were generated from Indian isolates of Toxoplasma gondii by transferring single tissue cysts from the brains of chronically infected mice to confluent murine macrophage (J774A.1) monolayers. The clones were then maintained continuously as tachyzoites in culture. Physical rupture of the tissue cysts and release of bradyzoites prior to seeding was found to be necessary for establishment of the parasite in culture. Although intact tissue cysts seeded over monolayers released bradyzoites spontaneously, they did not succeed in setting up an infection in the monolayers. Random amplified polymorphic DNA (RAPD)-PCR, which revealed distinct patterns for a clone and its progenitor, further confirmed the efficiency of the technique. The cloning technique was found to be simple and rapid compared to those involving limiting dilutions.

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Acknowledgements

The authors thank the Director and Joint Director (Academic), Indian Veterinary Research Institute, for providing the facilities to conduct the research. Thanks are also due to ICAR and IVRI for the fellowship awarded to the senior author. The authors are grateful to Dr. J.P. Dubey, USDA, for his constructive criticism on the manuscript and to Valsin Fournet, USDA, for assisting with the figures. The experiments comply with the current laws applicable to animal experimentation in India.

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Authors and Affiliations

  1. Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, Bareilly, 243 122, Uttar Pradesh, India

    C. Sreekumar, J. R. Rao, A. K. Mishra, D. Ray & G. C. Bansal

  2. ARS, ANRI, APDL, BARC, USDA, East Bldg. 1001, 10300 Baltimore Avenue, Beltsville, MD 20705-2350, USA

    C. Sreekumar

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  1. C. Sreekumar
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  2. J. R. Rao
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  3. A. K. Mishra
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  4. D. Ray
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  5. G. C. Bansal
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Correspondence to C. Sreekumar.

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Sreekumar, C., Rao, J.R., Mishra, A.K. et al. In vitro generation of cloned populations of Toxoplasma gondii . Parasitol Res 90, 489–492 (2003). https://doi.org/10.1007/s00436-003-0898-2

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  • DOI: https://doi.org/10.1007/s00436-003-0898-2

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