PURIFICATION AND CHARACTERIZATION OF <i>TOXOPLASMA GONDII</i> TACHYZOITE DNA

Johnson, Alan M.; Dubey, J. P.; Dame, John B.

doi:10.1038/icb.1986.37

Cited by 28 publications

(2 citation statements)

References 11 publications

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“…Genomic DNA was extracted from 10 9 tachyzoites by a routine protocol described previously by Johnson et al (1986). The DNA pellet was resuspended in 0 .…”

Section: Bradyzoite Preparation and Purificationmentioning

confidence: 99%

Toxoplasma gondiidense granule protein 3 (GRA3) is a type I transmembrane protein that possesses a cytoplasmic dilysine (KKXX) endoplasmic reticulum (ER) retrieval motif

et al. 2005

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Studies using antibodies to immunolocalize the Toxoplasma gondii dense granule protein GRA3, have shown that this protein associates strongly with the parasitophorous vacuole membrane (PVM). However, as there was no predicted membrane-spanning domain this highlighted an unanswered paradox. We demonstrate that the previously published sequence for GRA3 is actually an artificial chimera of 2 proteins. One protein, of molecular weight 65 kDa, shares the C-terminus with published GRA3 and possesses no significant sequence similarity with any protein thus far deposited in Genbank. The second, with a predicted molecular weight of 24 kDa shares the N-terminal region, is recognized by the monoclonal antibody 2H11 known to react with the dense granules of T. gondii and is therefore the authentic GRA3. The corrected GRA3 has an N-terminal secretory signal sequence and a transmembrane domain consistent with its insertion into the PVM. Antibodies to recombinant GRA3 recognize a protein of 24 kDa in T. gondii excretory-secretory antigen preparations. The signal peptide is necessary and sufficient to target GFP to the dense granules and parasitophorous vacuole. A homologue was identified in Neospora caninum. Finally, GRA3 possesses a dilysine 'KKXX' endoplasmic reticulum (ER) retrieval motif that rationalizes its association with PVM and possibly the host cell ER.

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“…Genomic DNA was extracted from 10 9 tachyzoites by a routine protocol described previously by Johnson et al (1986). The DNA pellet was resuspended in 0 .…”

Section: Bradyzoite Preparation and Purificationmentioning

confidence: 99%

Toxoplasma gondiidense granule protein 3 (GRA3) is a type I transmembrane protein that possesses a cytoplasmic dilysine (KKXX) endoplasmic reticulum (ER) retrieval motif

et al. 2005

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“…Recent advances in molecular-biology are likely to lead to the development of recombinant vaccines. Purification and characterisation of Toxoplasma DNA is currently being carried out in a number of laboratories with this long term aim in mind [7,36,37].…”

Section: Immunisationmentioning

confidence: 99%

Recent developments in the prevention and treatment of congenital toxoplasmosis

Dutton

1989

Int Ophthalmol

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The manifestations of congenital toxoplasmosis vary considerably in degree, characteristics and time of onset. Options for prevention of the disease include the appropriate disposal of cat litter and the avoidance of ingestion of both contaminated food and undercooked meat by pregnant women. Immunisation of the domestic cat population is a consideration for the future. Alternatively, immunisation of sero-negative pregnant women awaits the introduction of an effective and safe reagent. Current treatment modalities are not universally effective and new drugs are the subject of active development and research. Screening of pregnant women or perhaps newborn infants are potential options but the cost effectivity has yet to be established in many countries and the results of treatment during pregnancy and early childhood are encouraging but are as yet unproven. The majority of patients with congenital toxoplasmosis eventually develop toxoplasmic retinochoroiditis. Current treatment options for this condition are outlined. Acquired immune deficiency syndrome (AIDS) may, in some cases, represent a recrudescence of congenital intracerebral infection. Current treatment strategies for this condition are discussed.

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Enzymes for Technical Applications

et al. 2002

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Introduction Historical Outline Enzymes for the Detergent Industry Introduction History Overview of Enzymes Latest Innovations Future Perspectives Enzymes for the Starch Industry Introduction History Enzymes Latest Innovations Perspectives Enzymes for Biofuel Introduction History Enzymes and Latest Innovations Future Perspectives Enzymes for the Textile Industry Introduction History Enzymes Latest Innovations Enzymes for the Pulp and Paper Industry Introduction Overview of Selected Applications Enzymes Future Perspectives Enzymes for Organic Synthesis Introduction History Enzymes Latest Innovations Perspectives Enzymes for Processing of Fats and Oils Introduction History Enzymes Latest Innovations Perspectives Key Technologies for the Discovery of Industrial Enzymes Exploring Nature's Diversity Protein Optimization Conclusion Acknowledgments

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PURIFICATION AND CHARACTERIZATION OF TOXOPLASMA GONDII TACHYZOITE DNA

Cited by 28 publications

References 11 publications

Toxoplasma gondiidense granule protein 3 (GRA3) is a type I transmembrane protein that possesses a cytoplasmic dilysine (KKXX) endoplasmic reticulum (ER) retrieval motif

Toxoplasma gondiidense granule protein 3 (GRA3) is a type I transmembrane protein that possesses a cytoplasmic dilysine (KKXX) endoplasmic reticulum (ER) retrieval motif

Recent developments in the prevention and treatment of congenital toxoplasmosis

Enzymes for Technical Applications

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