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Original Article

Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii

Eui-Sun Son1, Kyoung-Ju Song2, Jong-Chul Shin2, Ho-Woo Nam1
The Korean Journal of Parasitology 2001;39(2):133-141.
Published online: June 30, 2001

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Corresponding author (howoo@cmc.cuk.ac.kr)
• Received: May 10, 2001   • Accepted: June 1, 2001

copyright © 2001 by The Korean Society for Parasitology

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Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii
Korean J Parasitol. 2001;39(2):133-141.   Published online June 30, 2001
DOI: https://doi.org/10.3347/kjp.2001.39.2.133
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Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii
Korean J Parasitol. 2001;39(2):133-141.   Published online June 30, 2001
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Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii
Image Image Image Image Image Image
Fig. 1 RACE and nested PCR products of peroxiredoxin from Toxoplasma gondii. The DNA fragments of lane 1 and 3 were amplified by 5'- and 3'-RACE, respectively. Lane 2 and 4 were nested PCR products amplified on the DNA of lane 1 and 3 as template, respectively.
Fig. 2 cDNA and deduced amino acid sequences of peroxiredoxin from Toxoplasma gondii. An open reading frame is written in bold. Primers used for 5'- and 3'-RACE and for recombinant fusion protein are indicated and labeled by arrows. Reactive cysteine residues are underlined at the position 51 and 171. Nucleotide sequence data are available in the GenBank™ under the accession number AF305718.
Fig. 3 Alignment of the deduced amino acid sequence of Toxoplasma gondii peroxiredoxin with those of 2-Cys peroxiredoxins from other organisms. Two reactive cysteine domains are characterized by asterisks (*). GenBank accession numbers for other organisms are as follow: Arabidopsis thaliana, Y10478; Burgia malayi Prx2, U47100; Escherichia coli AhpC, D13187; Entamoeba histolytica, X70996; Fasciola hepatica, U88577; Onchocerca volvulus Prx1, U31052; Rat Prx III, U06099; Rat Prx IV, D30035; Trypanosoma brucei, U26666; Burgia malayi Prx1, U34251; Dirofilaria immitis Prx1, AF004167; Leishmania major, AF069386; and Onchocerca volvulus Prx2, AF029247.
Fig. 4 Western blots to confirm the specificities of anti-Prx and -SOD antibodies. Each antibody was reacted exclusively with extracellular Toxoplasma gondii (RH) and intracellular T. gondii which infected into Vero cells for 24 hr and 48 hr, not with Vero cells only.
Fig. 5 Localization of peroxiredoxin in Toxoplasma gondii tachyzoites (A) and those within Vero cells (B). Rabbit anti-TgPrx antiserum was counter-stained with mouse monoclonal anti-dense granule protein antibody (Tg378) as a control as PVM secretion.
Fig. 6 A. Effect of artemisinin on Vero cells. Vero cells were treated with 1 µg/ml artemisinin. C1, untreated Vero cells only. B. Effect of artemisinin on the intracellular Toxoplasma gondii. Vero cells infected with T. gondii were treated with 1 µg/ml artemisinin. C1, untreated, uninfected Vero cells; and C2, untreated, infected Vero cells.

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Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii
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