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Establishment of transient and stable transfection systems for Babesia ovata

Overview of attention for article published in Parasites & Vectors, March 2016
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  • Above-average Attention Score compared to outputs of the same age (54th percentile)
  • Above-average Attention Score compared to outputs of the same age and source (52nd percentile)

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3 tweeters

Citations

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26 Dimensions

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41 Mendeley
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Title
Establishment of transient and stable transfection systems for Babesia ovata
Published in
Parasites & Vectors, March 2016
DOI 10.1186/s13071-016-1439-z
Pubmed ID
Authors

Hassan Hakimi, Junya Yamagishi, Yuto Kegawa, Osamu Kaneko, Shin-ichiro Kawazu, Masahito Asada

Abstract

Bovine 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. In 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. After 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. The current study provides a fundamental molecular tool to aid in molecular and cellular studies of B. ovata.

Twitter Demographics

The data shown below were collected from the profiles of 3 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 41 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 41 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 7 17%
Student > Master 6 15%
Student > Ph. D. Student 6 15%
Student > Bachelor 5 12%
Student > Doctoral Student 4 10%
Other 6 15%
Unknown 7 17%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 12 29%
Veterinary Science and Veterinary Medicine 6 15%
Agricultural and Biological Sciences 4 10%
Immunology and Microbiology 3 7%
Medicine and Dentistry 3 7%
Other 4 10%
Unknown 9 22%

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 07 April 2016.
All research outputs
#3,403,576
of 7,516,464 outputs
Outputs from Parasites & Vectors
#888
of 2,041 outputs
Outputs of similar age
#124,211
of 275,386 outputs
Outputs of similar age from Parasites & Vectors
#84
of 185 outputs
Altmetric has tracked 7,516,464 research outputs across all sources so far. This one has received more attention than most of these and is in the 54th percentile.
So far Altmetric has tracked 2,041 research outputs from this source. They receive a mean Attention Score of 3.6. This one has gotten more attention than average, scoring higher than 55% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 275,386 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 54% of its contemporaries.
We're also able to compare this research output to 185 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 52% of its contemporaries.