| Summary: | The development of genetic manipulation techniques has been reported in many protozoan parasites over the past few years. However, these techniques have not been established for <i>Babesia microti</i>. Here, we report the first successful transient transfection of <i>B. microti</i>. The plasmids containing the firefly luciferase reporter gene were transfected into <i>B. microti</i> by an AMAXA 4D Nucleofection system. Twenty-four-hour synchronization, the <i>5′-actin</i> promoter, program FA100, and 50 μg of plasmid DNA constituted the best conditions for the transient transfection of <i>B. microti</i>. This finding is the first step towards a stable transfection method for <i>B. microti</i>, which may contribute to a better understanding of the biology of the parasite.
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