Establishment of a transient transfection system for Babesia sp. Xinjiang with the use of homologous promoters

作者：Xiaoxing Wang, Jinming Wang, Junlong Liu, Jifei Yang, Zhaoyong Lv, Aihong Liu, Youquan Li, Yubin Li, Haiou Lan, Guangyuan Liu, Jian-Xun Luo, Guiquan Guan, Hong Yin · 发表于：Research Square · 年份：2020 · DOI：10.21203/rs.3.rs-90736/v1 · 研究领域：Vector-Borne Animal Diseases、Insect Resistance and Genetics、Vector-borne infectious diseases

Abstract Background 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 ( Bxj ), which is the causative pathogen of ovine babesiosis mainly in China, the efficiency of these techniques remain unclear. MethodsFirst, a plasmid bearing the elongation factor-1 alpha promoter, as well as the firefly luciferase reporter gene and rap stop region were transfected into Bxj by electroporation and nucleoporation to determine the most suitable transfection solution. Then, six program setting were evaluated to confirm the best for Bxj transient transfection and a series of different amounts of plasmid DNA were applied to generate relatively high luminescence values. Finally, the activities of four promoters derived from Bxj were evaluated using the developed transient transfection system. ConclusionsIn this study, a transient transfection system for Bxj was optimized. These findings provided critical information for Bxj genetic manipulation, as an essential tool to identify virulence factors and to further elucidate the basic biology of pathogens.