Stable expression of green fluorescent protein and targeted disruption of thioredoxin peroxidase-1 gene in Babesia bovis with the WR99210/dhfr selection system

Asada, Masahito; Tanaka, Miho; Goto, Yasuyuki; Yokoyama, Naoki; Inoue, Noboru; Kawazu, Shin-ichiro

doi:10.1016/j.molbiopara.2011.11.001

Cited by 37 publications

(70 citation statements)

References 33 publications

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“…GFP-fluorescent B. bovis populations were established previously [13]. Briefly, the GFP-expressing plasmid was constructed.…”

Section: Methodsmentioning

confidence: 99%

“…Here we have observed gliding motility using time-lapse video microscopy of GFP-expressing B. bovis merozoites that were developed in our previous study [13]. Time-lapse video images delineated the sequential process of parasite-infected RBC (IRBC) rupture, merozoite egress from IRBCs, gliding motility of the merozoites, attachment and invasion of merozoites into new RBCs, and formation and breakdown of PVs.…”

Section: Introductionmentioning

confidence: 96%

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Gliding Motility of Babesia bovis Merozoites Visualized by Time-Lapse Video Microscopy

et al. 2012

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Background Babesia bovis is an apicomplexan intraerythrocytic protozoan parasite that induces babesiosis in cattle after transmission by ticks. During specific stages of the apicomplexan parasite lifecycle, such as the sporozoites of Plasmodium falciparum and tachyzoites of Toxoplasma gondii, host cells are targeted for invasion using a unique, active process termed “gliding motility”. However, it is not thoroughly understood how the merozoites of B. bovis target and invade host red blood cells (RBCs), and gliding motility has so far not been observed in the parasite.Methodology/Principal FindingsGliding motility of B. bovis merozoites was revealed by time-lapse video microscopy. The recorded images revealed that the process included egress of the merozoites from the infected RBC, gliding motility, and subsequent invasion into new RBCs. The gliding motility of B. bovis merozoites was similar to the helical gliding of Toxoplasma tachyzoites. The trails left by the merozoites were detected by indirect immunofluorescence assay using antiserum against B. bovis merozoite surface antigen 1. Inhibition of gliding motility by actin filament polymerization or depolymerization indicated that the gliding motility was driven by actomyosin dependent process. In addition, we revealed the timing of breakdown of the parasitophorous vacuole. Time-lapse image analysis of membrane-stained bovine RBCs showed formation and breakdown of the parasitophorous vacuole within ten minutes of invasion.Conclusions/SignificanceThis is the first report of the gliding motility of B. bovis. Since merozoites of Plasmodium parasites do not glide on a substrate, the gliding motility of B. bovis merozoites is a notable finding.

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“…GFP-fluorescent B. bovis populations were established previously [13]. Briefly, the GFP-expressing plasmid was constructed.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 96%

Gliding Motility of Babesia bovis Merozoites Visualized by Time-Lapse Video Microscopy

et al. 2012

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“…However, to serve as efficient live vectors for vaccination, recombinant attenuated B. bovis vaccines would be required to establish mild acute infection leading to persistence, express exogenous antigens throughout infection, and remain genetically stable. Several laboratories, including our own, have been able to stably transfect B. bovis [5][6][7], removing a technical barrier for creating recombinant marker and multivalent attenuated vaccines. However, previous work has not demonstrated the requisite in vivo features of such vaccines in cattle, the natural host of B. bovis.…”

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confidence: 99%

Acute and persistent infection by a transfected Mo7 strain of Babesia bovis

Suárez

Laughery

Schneider

et al. 2012

Molecular and Biochemical Parasitology

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“…This cytoplasmic expression pattern was also shown in typical 2-Cys peroxiredoxins of P. falciparum (PfTPx-1) [31], P. vivax (PvTPx-1) [11] and B. bovis (BbTPx-1) [27]. Recently, our group showed that BbTpx-1 gene disruption does not affect in vitro intraerythrocytic growth of B. bovis [2], as previously reported for the gene disrupted in P. berghei [30], indicating that the TPx-1 gene is not essential for the erythrocytic stage of Babesia parasites. In fact, Babesia parasites have other antioxidant proteins, such as catalase and Gpx [4, 8].…”

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confidence: 81%

Cloning and Characterization of a 2-Cys Peroxiredoxin from <i>Babesia gibsoni</i>

Masatani

Asada

Ichikawa‐Seki

et al. 2014

The Journal of Veterinary Medical Science

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Peroxiredoxins (Prxs) are a family of antioxidant enzymes. Here, we cloned a 2-Cys Prx, BgTPx-1, from the canine Babesia parasite B. gibsoni. Sequence identity between BgTPx-1 and 2-Cys Prx of B. bovis was 81% at the amino acid level. Enzyme activity assay by using recombinant BgTPx-1 (rBgTPx-1) indicated that BgTPx-1 has antioxidant activity. Antiserum from a mouse immunized with rBgTPx-1 reacted with parasite lysates and detect a protein with a monomeric size of 22 kDa and also a 44 kDa protein, which might be an inefficiently reduced dimer. BgTPx-1 was expressed in the cytoplasm of B. gibsoni merozoites. These results suggest that the BgTPx-1 may play a role to control redox balance in the cytoplasm of B. gibsoni.

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Stable expression of green fluorescent protein and targeted disruption of thioredoxin peroxidase-1 gene in Babesia bovis with the WR99210/dhfr selection system

Cited by 37 publications

References 33 publications

Gliding Motility of Babesia bovis Merozoites Visualized by Time-Lapse Video Microscopy

Gliding Motility of Babesia bovis Merozoites Visualized by Time-Lapse Video Microscopy

Acute and persistent infection by a transfected Mo7 strain of Babesia bovis

Cloning and Characterization of a 2-Cys Peroxiredoxin from <i>Babesia gibsoni</i>

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