Stable transformation of fluorescent proteins into Nosema bombycis by electroporation

Parasites & Vectors

Cheng Lu¹  Peng Chen¹  Qin Wu¹  Boyuan Deng¹  Minhui Pan¹  Na Gao¹  Congwu Hu¹  Xuhua Huang¹  Zhanqi Dong¹ 

[1] State Key Laboratory of Silkworm Genome Biology, Southwest University;

关键词: Microsporidia;    Bombyx mori;    Nosema bombycis;    Electro-transformation;   

DOI  :  10.1186/s13071-022-05236-4

来源: DOAJ

【 摘 要 】

Abstract Background Microsporidia are a group of intracellular parasitic eukaryotes, serious pathogens that cause widespread infection in humans, vertebrates, and invertebrates. Because microsporidia have a thick spore wall structure, the in vitro transformation, cell culture, and genetic operation technology of microsporidia are far behind that of other parasites. Methods In this study, according to an analysis of the life-cycle of microsporidia, Nosema bombycis, and different electro-transformation conditions, the transduction efficiency of introducing foreign genes into N. bombycis was systematically determined. Results We analyzed the direct electro-transformation of foreign genes into germinating N. bombycis using reporters under the regulation of different characteristic promoters. Furthermore, we systematically determined the efficiency of electro-transformation into N. bombycis under different electro-transformation conditions and different developmental stages through an analysis of the whole life-cycle of N. bombycis. These results revealed that foreign genes could be effectively introduced through a perforation voltage of 100 V pulsed for 15 ms during the period of N. bombycis sporeplasm proliferation. Conclusions We present an effective method for electro-transformation of a plasmid encoding a fluorescent protein into N. bombycis, which provides new insight for establishing genetic modifications and potential applications in these intracellular parasites. Graphical Abstract

【 授权许可】

Unknown