摘要：Theileria equi, an intraerythrocytic protozoan parasite, causes equine piroplasmosis, a disease which negatively impacts the global horse industry. Genetic manipulation is one of the research tools under development as a control method for protozoan parasites, but this technique needs to be established for
T.
equi. Herein, we report on the first development of a stable transgenic
T.
equi line expressing enhanced green fluorescent protein/blasticidin S deaminase (eGFP/BSD). To express the exogenous fusion gene in
T.
equi, regulatory regions of the
elongation factor-1 alpha (
ef-1α) gene were identified in
T. equi. An eGFP/BSD-expression cassette containing the
ef-1α gene promoter and terminator regions was constructed and integrated into the
T.
equi genome. On day 9 post-transfection, blasticidin-resistant
T.
equi emerged. In the clonal line of
T. equi obtained by limiting dilution, integration of the eGFP/BSD-expression cassette was confirmed in the designated B
-locus of the
ef-1α gene via PCR and Southern blot analyses
. Parasitaemia dynamics between the transgenic and parental
T.
equi lines were comparable in vitro. The eGFP/BSD-expressing transgenic
T.
equi and the methodology used to generate it offer new opportunities for better understanding of
T.
equi biology, with the add-on possibility of discovering effective control methods against equine piroplasmosis.
