Reporter gene expression in cell culture stages and oocysts of Eimeria nieschulzi (Coccidia, Apicomplexa).
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 beta-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 beta-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.
Kurth
Michael
M
Section Molecular Biotechnology, Technische Universität Dresden, 01062 Dresden, Germany. kurth.michael@googlemail.com
Entzeroth
Rolf
R
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis*; D003048:Coccidiosis / Q000469:parasitology; D004539:Eimeria / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000096:biosynthesis*; D033661:Oocysts / Q000378:metabolism; D051381:Rats; D014170:Transformation, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis*
D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D001616:beta-Galactosidase
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