Parasitology Research

, Volume 104, Issue 2, pp 303–310 | Cite as

Reporter gene expression in cell culture stages and oocysts of Eimeria nieschulzi (Coccidia, Apicomplexa)

Original Paper
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Abstract

The rat parasite Eimeria nieschulzi is a suitable model for transfection studies and was used as an additional model organism for the genus Eimeria. We describe the transfection of this apicomplexan parasites and the cultivation of transformed stages in cell culture and in vivo. The β-galactosidase or yellow fluorescent protein was expressed in all parasitic stages up to the second merozoite generation in vitro under control of the heterologous promoter region of Eimeria tenella mic1 gene previously described for E. tenella transfection. Pyrimethamine resistant E. nieschulzi parasites were obtained in vitro after transfection with a plasmid encoding the Toxoplasma gondii dhfr/ts-m2m3 gene. Co-transfection experiments with an YFP-plasmid resulted in pyrimethamine resistant and fluorescent parasitic stages. Infection of rats with transfected E. nieschulzi sporozoites directed to expression of β-galactosidase or YFP in oocysts. Co-transfection with YFP/DHFR-TS allowed selection of resistant parasites in vivo. Excreted transgenic oocysts showed arrangement of YFP expression which lead to questions about meiotic recombination frequency and mechanisms.

Keywords

Yellow Fluorescent Protein Pyrimethamine IEC6 Cell Coccidiosis Eimeria Species 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Notes

Acknowledgements

We appreciate and thank for financial support from the German Research Foundation (DFG). Grateful thanks go to Fiona Tomley and Julie Clark (Institute for Animal Health, Compton, UK) providing us with the plasmids and for the great support and successful collaboration. In this combination we thank the European Union for financing a research visit during the COST857 program. Many thanks go to Boris Striepen (University of Georgia, Athens, GA, USA) and Marc-Jan Gubbels (now Boston College, USA) for providing lab capacities and mentoring during a DFG founded research period. We also thank Bill Chobotar (Andrews University, USA) for reading and correcting the manuscript.

Declaration

We declare that the experiments comply with the current laws of Germany where they were performed.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Section Molecular BiotechnologyTechnische Universität DresdenDresdenGermany
  2. 2.Institute of ZoologyTechnische Universität DresdenDresdenGermany

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