Dataset Information

Establishment of a stable transfection system for genetic manipulation of Babesia gibsoni.

ABSTRACT: BACKGROUND:Genetic manipulation techniques, such as transfection, have been previously reported in many protozoan parasites. In Babesia, stable transfection systems have only been established for bovine Babesia parasites. We recently reported a transient transfection system and the selection of promoter candidates for Babesia gibsoni. The establishment of a stable transfection system for B. gibsoni is considered to be urgent to improve our understanding of the basic biology of canine Babesia parasites for a better control of babesiosis. RESULTS:GFP-expressing parasites were observed by fluorescence microscopy as early as two weeks after drug selection, and consistently expressed GFP for more than 3 months without drug pressure. Genome integration was confirmed by PCR, sequencing and Southern blot analysis. CONCLUSIONS:We present the first successful establishment of a stable transfection system for B. gibsoni. This finding will facilitate functional analysis of Babesia genomes using genetic manipulation and will serve as a foundation for the development of tick-Babesia and host-Babesia infection models.

SUBMITTER: Liu M

PROVIDER: S-EPMC5914073 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Establishment of a stable transfection system for genetic manipulation of Babesia gibsoni.

Liu Mingming M Adjou Moumouni Paul Franck PF Asada Masahito M Hakimi Hassan H Masatani Tatsunori T Vudriko Patrick P Lee Seung-Hun SH Kawazu Shin-Ichiro SI Yamagishi Junya J Xuan Xuenan X

Parasites & vectors 20180423 1

<h4>Background</h4>Genetic manipulation techniques, such as transfection, have been previously reported in many protozoan parasites. In Babesia, stable transfection systems have only been established for bovine Babesia parasites. We recently reported a transient transfection system and the selection of promoter candidates for Babesia gibsoni. The establishment of a stable transfection system for B. gibsoni is considered to be urgent to improve our understanding of the basic biology of canine Bab ...[more]

PMID: 29685172

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Project description:BackgroundSeveral members of genus Babesia are important pathogens causing babesiosis in dogs. In China, at least five Babesia species have been described in dogs or ticks. This study sought to determine the prevalence and molecular characteristics of various Babesia spp. in dogs in cities in Shaanxi Province in China, including Xi'an and Hanzhong.MethodsA total of 371 blood samples were collected from pet dogs presenting to veterinary clinics in the cities of Xi'an and Hanzhong in Shaanxi, China. Babesia spp. DNA was detected via amplification of partial 18S rRNA genes by semi-nested PCR. Almost full-length 18S rRNA, ITS, partial TRAP and complete cytb genes were recovered for analysis of the genetic characteristics and relationships with known isolates.ResultsA single species, Babesia gibsoni, was identified in dogs in Xi'an and Hanzhong. Consistently, B. gibsoni was also detected in 14 ticks collected from positive dogs. Sequence similarities and phylogenetic analysis suggested that the isolates identified herein showed a closer genetic relationship with isolates from East Asian countries rather than India, Bangladesh, or the USA. Sequence analysis based on tandem repeat analysis of the TRAP gene further revealed that specific haplotypes were circulating in both Xi'an and Hanzhong, with no specific regionality. In addition, 10.9% of all isolates with atovaquone (ATV)-resistance were identified because of M121I mutation in the deduced cytb protein.ConclusionsThis study revealed a high prevalence rate of Babesia infection. Babesia gibsoni was the only Babesia species identified in cases of canine babesiosis in the cities of Xi'an and Hanzhong cities in Shaanxi, China. In addition, the TRAP gene presented high genetic diversity across isolates. Such information is useful for elucidating the epidemiological characteristics of canine babesiosis, as well as the overall genetic diversity of Babesia spp. circulating in dog populations in Shaanxi Province.

Dataset Information

Establishment of a stable transfection system for genetic manipulation of Babesia gibsoni.

Publications

Establishment of a stable transfection system for genetic manipulation of Babesia gibsoni.

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