Transient transfection of Babesia ovis using heterologous promoters
Transient transfection of Babesia ovis using heterologous promoters
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DOI：
10.1016/j.ttbdis.2019.101279
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发表时间：
2019-10-01
影响因子：
3.2
通讯作者：
Antunes, Sandra
中科院分区：
文献类型：
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作者：
Rosa, Catarina;Asada, Masahito;Antunes, Sandra
巴贝斯虫是一种公认的新兴蜱传疾病，巴贝斯虫病的病原体，是一个重要的动物和人类健康问题，具有世界范围的社会经济影响。基因操作技术的发展，如转染技术，是关键的，以提高知识，这些研究不足的寄生虫的生物学更好的疾病控制策略。对于引起绵羊巴贝斯虫病（一种由蜱传播的小型反刍动物疾病）的巴贝斯虫来说，这些工具还不可用。本研究是基于存在可互换的跨物种的功能性启动子之间的Bababellus物种。在此，我们首次描述了B。ovis瞬时转染，使用两个异源启动子，来自B的ef-1 α-B基因间区。bovis和B.卵形的它们驱动报告荧光素酶在B中表达的能力。ovis支持它们的跨物种功能。此外，ef-1 α-B启动子区来自B.与对照相比，卵圆形导致统计学上显著更高的发光值，因此可能是用于稳定基因表达的合适启动子。使用qPCR对转染效率的评估表明，较高的发光水平是由于启动子强度而不是较高的转染效率。这些发现代表了B方法发展的一个进步。ovis基因操作无疑是研究该寄生虫基础生物学的必要工具，包括其生活史、寄生虫与宿主细胞的相互作用和毒力因子。
Babesia species, etiological agents of babesiosis, a recognized emerging tick-borne disease, are a significant animal and human health concern with a worldwide socio-economic impact. The development of genetic manipulation techniques, such as transfection technology, is pivotal to improve knowledge regarding the biology of these poorly studied parasites towards better disease control strategies. For Babesia ovis, responsible for ovine babesiosis, a tick-borne disease of small ruminants, these tools are not yet available. The present study was based on the existence of interchangeable cross-species functional promoters between Babesia species. Herein, we describe for the first time B. ovis transient transfection using two heterologous promoters, the ef-1 alpha-B intergenic regions from B. bovis and B. ovata. Their ability to drive expression of a reporter luciferase in B. ovis supports their cross-species functionality. Also, the ef-1 alpha-B promoter region from B. ovata resulted in statistically significantly higher luminescence values in comparison to the control, thus a possibly suitable promoter for stable gene expression. Evaluation of transfection efficiency using qPCR demonstrated that higher luminescence levels were due to promoter strength rather than a higher transfection efficiency. These findings represent a step forward in the development of methods for B. ovis genetic manipulation, an undoubtedly necessary tool to study this parasite basic biology, including its life cycle, the parasite interactions with host cells and virulence factors.