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- W2803165744 abstract "Abstract Babesia bigemina and Babesia bovis , are the two major causes of bovine babesiosis, a global neglected disease in need of improved methods of control. Here, we describe a shared method for the stable transfection of these two parasites using electroporation and blasticidin/blasticidin deaminase as a selectable marker. Stably transfected B . bigemina and B . bovis were obtained using a common transfection plasmid targeting the enhanced green fluorescent protein-BSD ( egfp-bsd ) fusion gene into the elongation factor-1α ( ef-1α ) locus of B . bigemina and B . bovis under the control of the B . bigemina ef-1α promoter. Sequencing, Southern blotting, immunoblotting and immunofluorescence analysis of parasite-infected red blood cells, demonstrated that the egfp-bsd gene was expressed and stably integrated solely into the ef-1α locus of both, B . bigemina and B . bovis . Interestingly, heterologous B . bigemina ef-1α sequences were able to drive integration into the B . bovis genome by homologous recombination, and the stably integrated B . bigemina ef-1α -A promoter is fully functional in B . bovis . Collectively, the data provides a new tool for genetic analysis of these parasites, and suggests that the development of vaccine platform delivery systems based on transfected B . bovis and B . bigemina parasites using homologous and heterologous promoters is feasible." @default.
- W2803165744 created "2018-06-01" @default.
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- W2803165744 date "2018-04-17" @default.
- W2803165744 modified "2023-10-14" @default.
- W2803165744 title "Stable transformation of Babesia bigemina and Babesia bovis using a single transfection plasmid" @default.
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- W2803165744 doi "https://doi.org/10.1038/s41598-018-23010-4" @default.
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