Molecular genetic transfection of the coccidian parasite Sarcocystis neurona

Rajshekhar Y. Gaji, Deqing Zhang, Cormac C. Breathnach, Shipra Vaishnava, Boris Striepen, Daniel K. Howe

Research output: Contribution to journalArticlepeer-review

22 Scopus citations


Sarcocystis neurona is an apicomplexan parasite that is the major cause of equine protozoal myeloencephalitis (EPM). The biology of this pathogen remains poorly understood in part due to unavailability of molecular genetic tools. Hence, with an objective to develop DNA transfection capabilities for S. neurona, the 5′ flanking region of the SnSAG1 gene was isolated from a genomic library and used to construct expression plasmids. In transient assays, the reporter molecules β-galactosidase (β-gal) and yellow fluorescent protein (YFP) could be detected in electroporated S. neurona, thereby confirming the feasibility of transgene expression in this organism. Stable transformation of S. neurona was achieved using a mutant dihydrofolate reductase thymidylate synthase (DHFR-TS) gene of Toxoplasma gondii that confers resistance to pyrimethamine. This selection system was used to create transgenic S. neurona that stably express β-gal and YFP. As shown in this study, these transgenic clones can be useful for analyzing growth rate of parasites in vitro and for assessing drug sensitivities. More importantly, the DNA transfection methods described herein should greatly facilitate studies examining intracellular parasitism by this important coccidian pathogen.

Original languageEnglish
Pages (from-to)1-9
Number of pages9
JournalMolecular and Biochemical Parasitology
Issue number1
StatePublished - Nov 2006

Bibliographical note

Funding Information:
We gratefully acknowledge the technical assistance of Dr. David Horohov. This research was supported by grants from the Amerman Family Foundation and Fort Dodge Animal Health (to DKH) and grants from NIH-NIAID (to BS). R. Gaji is supported by a Paul Mellon Graduate Student Fellowship in Equine Veterinary Science. Published as Kentucky Agricultural Experiment Station Article No. 06-14-020.


  • Apicomplexa
  • Coccidia
  • Fluorescence activated cell sorting
  • Sarcocystis neurona
  • Stable transformation
  • Transfection

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology


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