Establishment of a transient transfection system for Babesia sp. Xinjiang using homologous promoters

文献类型： 外文期刊

第一作者： Wang, Xiaoxing

作者： Wang, Xiaoxing;Wang, Jinming;Liu, Junlong;Yang, Jifei;Lv, Zhaoyong;Liu, Aihong;Li, Youquan;Li, Yubin;Lan, Haiou;Liu, Guangyuan;Luo, Jianxun;Guan, Guiquan;Yin, Hong;Yin, Hong

作者机构：

关键词： Babesia sp; Xinjiang; Transient transfection; Promoter activity; Genetic manipulation

期刊名称：PARASITOLOGY RESEARCH （ 影响因子：2.289； 五年影响因子：2.403 ）

ISSN： 0932-0113

年卷期： 2021 年 120 卷 10 期

页码：

收录情况： SCI

摘要： Babesia species, the agentic pathogens of human and animal babesiosis, are spread worldwide. Over the last decade, genetic manipulation approaches have been applied with many protozoan parasites, including Plasmodium falciparum, Trypanosoma cruzi, Cryptosporidium parvum, Theileria annulata, Theileria parva, Babesia bovis, Babesia bigemina, Babesia ovata, Babesia gibsoni, and Babesia ovis. For Babesia sp. Xinjiang (BspXJ), which is the causative pathogen of ovine babesiosis mainly in China, the efficiency of these techniques remains unclear. Firstly, a plasmid bearing the elongation factor-1 alpha promoter and the firefly luciferase reporter gene and rap stop region were transfected into BspXJ by electroporation and nucleoporation to determine the most suitable transfection solution. Then, six program settings were evaluated to confirm the best for BspXJ transient transfection, and a series of different amounts of plasmid DNA were transfected to generate relatively high luminescence values. Finally, the activities of four promoters derived from BspXJ were evaluated using the developed transient transfection system. After evaluating of various transfection parameters, the human T cell nucleofector solution, program V-024 and 20 mu g of plasmid DNA were selected as the most favorable conditions for BspXJ transient transfection. These findings provide critical information for BspXJ genetic manipulation, an essential tool to identify virulence factors and to further elucidate the basic biology of this parasite.

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