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Abstract Objective: To construct Toxoplasma gondii(T. gondii) mutant for expression of green fluorescent protein (GFP) in apicoplast and establish the simple methods for apicoplast detection. Methods: Acyl carrier protein (ACP) gene was amplified from T. gondii genomic DNA and used to construct the transfection plasmid pHX-ACP-GFP, which was transfected into T. gondii tachyzoites. The mutant was selected by mycophenolic acid and xanthine. The expression of GFP was detected by confocal microscope and Western blotting. Results: The mutant was selected by mycophenolic acid and xanthine for one week. The green fluorescent aggregated in apicoplast was observed by confocal microscope. The transit peptide and mature ACPs were detected using anti-GFP tag antibody by Western blotting. Conclusion: Apicoplast could be labeled by ACP-GFP fusion protein which was detected by antiGFP tag antibody.
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