Stable transfection system for Babesia sp. Xinjiang.

Area of Science:

• Veterinary Parasitology
• Molecular Biology
• Protozoology

Background:

• Stable transfection systems are established for various protozoan parasites.
• Babesia sp. Xinjiang (Bxj) causes ovine babesiosis in China, lacking such a system.
• Genetic manipulation is crucial for understanding parasite biology and identifying drug targets.

Purpose of the Study:

• To develop a stable transfection system for Babesia sp. Xinjiang (Bxj).
• To enable genetic manipulation for enhanced research into Bxj biology.

Main Methods:

• Evaluated blasticidin (BSD) and WR99210 for inhibition efficiency in Bxj.
• Constructed a plasmid with BSD selectable marker, green fluorescent protein (GFP), and rap 3' TR.
• Integrated the plasmid into the Bxj ef-1α site via homologous recombination and transfected Bxj merozoites.

Main Results:

• Successfully generated stable transfected parasite lines (Bxj-c1, Bxj-c2, Bxj-c3).
• Confirmed exogenous gene integration into the Bxj genome using PCR and sequencing.
• Demonstrated sustained expression of GFP-BSD for 11 months via Western blot and IFA.

Conclusions:

• A stable transfection system for Babesia sp. Xinjiang (Bxj) has been successfully established.
• This platform is expected to significantly advance basic research on Bxj.