ArticleParasites & vectors2016
Establishment of transient and stable transfection systems for Babesia ovata.
Article in Parasites & vectors, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 33 citations in OpenAlex.
- Babesia duncani, a Model Organism for Investigating Intraerythrocytic Parasitism and Novel Antiparasitic Therapeutic Strategies.The Journal of infectious diseases · 2024Review
- Stable expression of red fluorescent protein-blasticidin deaminase fusion gene (rfp-bsd) as a selectable marker for DNA transfection in Babesia ovata.The Journal of veterinary medical science · 2024Article
- Transfection ofBio-protocol · 2024Article
- Establishment of a stable transfection and gene targeting system inFrontiers in cellular and infection microbiology · 2023Article
- Advances in understanding red blood cell modifications by Babesia.PLoS pathogens · 2022Review
- Establishment of a Transient and Stable Transfection System forFrontiers in cellular and infection microbiology · 2022Article
- Recent Advances in Molecular Genetic Tools forVeterinary sciences · 2021Review
- Establishment of a transient transfection system for Babesia sp. Xinjiang using homologous promoters.Parasitology research · 2021Article
- Stable transfection system for Babesia sp. Xinjiang.Parasites & vectors · 2021Article
- Molecular Survey ofVeterinary sciences · 2021Article
- A framework for signaling throughout the life cycle of Babesia species.Molecular microbiology · 2021Review
- Development of a stable transgenic Theileria equi parasite expressing an enhanced green fluorescent protein/blasticidin S deaminase.Scientific reports · 2021Article
- Establishment of a stable transfection method in Babesia microti and identification of a novel bidirectional promoter of Babesia microti.Scientific reports · 2020Article
- Transient Transfection of the Zoonotic ParasitePathogens (Basel, Switzerland) · 2020Article
- High resolution melting analysis of the 18S rRNA gene for the rapid diagnosis of bovine babesiosis.Parasites & vectors · 2019Article
- InitiatedPathogens (Basel, Switzerland) · 2019Article
- Genome Editing ofmSphere · 2019Article
- Knockout of Babesia bovis rad51 ortholog and its complementation by expression from the BbACc3 artificial chromosome platform.PloS one · 2019Article
- Establishment of a stable transfection system for genetic manipulation of Babesia gibsoni.Parasites & vectors · 2018Article
- Whole-genome assembly of Babesia ovata and comparative genomics between closely related pathogens.BMC genomics · 2017Article
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Authors and funding
6 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundBovine babesiosis is a tick-borne disease caused by several species of Babesia which produce acute and fatal disease in cattle and affect livestock industry worldwide. Babesia ovata is a benign species widespread in east Asian countries and causes anemia, particularly in cattle which are co-infected with Theileria orientalis. The development of genetic manipulation methods is necessary to improve our understanding of the basic biology of protozoan pathogens toward a better control of disease. Such tools have not been developed for B. ovata, and are the aim of this study.
methodsIn this study we transfected constructs that were designed to evaluate the ability of several B. ovata promoter candidates to drive expression of a reporter luciferase. We found that the elongation factor-1 alpha intergenic region (ef-1α IG) and the actin 5' non-coding region (NR) had highest promoter activities. To establish a stable transfection system, we generated a plasmid construct in which the ef-1α IG promoter drives gfp expression, and the actin 5' NR mediates expression of the selectable marker hdhfr. The plasmid was designed for episomal transfection, as well as to integrate by double cross-over homologous recombination into the ef-1α locus. Circular or linearized plasmid was transfected by electroporation into in vitro cultured B. ovata and retention of the plasmid was facilitated by drug selection with 5 nM WR99210 initiated 48 h after transfection.
resultsAfter one-week cultivation with WR99210, GFP-expressing parasites were observed by fluorescence microscopy. Integration of the plasmid construct into the ef-1α locus was confirmed by PCR, Southern blot analysis, and sequencing of recombination sites. These results confirm successful development of a stable transfection system for B. ovata.
conclusionThe current study provides a fundamental molecular tool to aid in molecular and cellular studies of B. ovata.
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