Transient transfection of intraerythrocytic Babesia gibsoni using elongation factor-1 alpha promoter

Publication date: Available online 17 July 2017 Source:Molecular and Biochemical Parasitology Author(s): Mingming Liu, Masahito Asada, Shinuo Cao, Paul Franck Adjou Moumouni, Patrick Vudriko, Artemis Efstratiou, Hassan Hakimi, Tatsunori Masatani, Fujiko Sunaga, Shin-ichiro Kawazu, Junya Yamagishi, Xuenan Xuan The development of gene manipulation techniques has been reported in many protozoan parasites over the past few years. However, these techniques have not yet been established for Babesia gibsoni. Here, we report for the first time, the successful transient transfection of B. gibsoni. The plasmid containing the firefly luciferase reporter gene (pBS-ELA) was transfected into B. gibsoni by an AMAXA 4D Nucleofector™ device. Transfection using program FA113 and Lonza buffer SF showed the highest luciferase expression. Twenty micrograms of plasmid produced the highest relative transfection efficiency. The fluorescent protein-expressing parasites were determined by GFP-containing plasmid (pBS-EGA) at 48 and 72hours post transfection. This finding is the first step towards a stable transfection method for B. gibsoni, which may contribute to a better understanding of the biology of the parasite. Graphical abstract
Source: Molecular and Biochemical Parasitology - Category: Parasitology Source Type: research