Disruption of the Toxoplasma gondii bradyzoite-specific gene BAG1 decreases in vivo cyst formation

Yi Wei Zhang, Kami Kim, Yan Fen Ma, Murray Wittner, Herbert B. Tanowitz, Louis M. Weiss

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Abstract

The bradyzoite stage of the Apicomplexan protozoan parasite Toxoplasma gondii plays a critical role in maintenance of latent infection. We reported previously the cloning of a bradyzoite-specific gene BAG1/hsp30 (previously referred to as BAG5) encoding a cytoplasmic antigen related to small heat shock proteins. We have now disrupted BAG1 in the T. gondii PLK strain by homologous recombination. H7, a cloned null mutant, and Y8, a control positive for both cat and BAG1, were chosen for further characterization. Immunofluorescence and Western blot analysis of bradyzoites with BAG1 antisera demonstrated expression of BAG1 in the Y8 and the PLK strain but no expression in H7. All three strains expressed a 116 kDa bradyzoite cyst wall antigen, a 29 kDa matrix antigen and the 65 kDa matrix reactive antigen MAG1. Mice inoculated with H7 parasites formed significantly fewer cysts than those inoculated with the Y8 and the PLK strains. H7 parasites were complemented with BAG1 using phleomycin selection., Cyst formation in vivo for the BAG1-complemented H7 parasites was similar to wild-type parasites. We therefore conclude that BAG1 is not essential for cyst formation, but facilitates formation of cysts in vivo.

Original languageEnglish (US)
Pages (from-to)691-701
Number of pages11
JournalMolecular Microbiology
Volume31
Issue number2
DOIs
StatePublished - 1999

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Toxoplasma
Cysts
Parasites
Antigens
Genes
Phleomycins
Small Heat-Shock Proteins
Homologous Recombination
Fluorescent Antibody Technique
Organism Cloning
Immune Sera
Cats
Western Blotting
Maintenance
Infection

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Disruption of the Toxoplasma gondii bradyzoite-specific gene BAG1 decreases in vivo cyst formation. / Zhang, Yi Wei; Kim, Kami; Ma, Yan Fen; Wittner, Murray; Tanowitz, Herbert B.; Weiss, Louis M.

In: Molecular Microbiology, Vol. 31, No. 2, 1999, p. 691-701.

Research output: Contribution to journalArticle

Zhang, Yi Wei ; Kim, Kami ; Ma, Yan Fen ; Wittner, Murray ; Tanowitz, Herbert B. ; Weiss, Louis M. / Disruption of the Toxoplasma gondii bradyzoite-specific gene BAG1 decreases in vivo cyst formation. In: Molecular Microbiology. 1999 ; Vol. 31, No. 2. pp. 691-701.
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