Characterization and in vitro translation of Toxoplasma gondii ribonucleic acid |
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| Affiliation: | 1. Department of Medicine, College of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan;2. Division of Chest Medicine, Department of Internal Medicine, Shin Kong Wu Ho-Su Memorial Hospital;3. Department of Urology, Taipei Medical University-Shuang Ho Hospital, New Taipei City, Taiwan;4. Department of Urology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan;5. Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan;6. Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan;7. Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan;8. Department of Medical Research, Shuang Ho Hospital, Taipei Medical University, New Taipei City, Taiwan;1. The Rockefeller University, 1230 York Avenue, New York, NY10021, USA;2. School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK;3. Institute of Parasitology, Biology Centre, Czech Academy of Sciences, 37005 Ceske Budejovice, Czech Republic |
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| Abstract: | RNA was extracted from purified tachyzoites of Toxoplasma gondii (RH strain) by sequential centrifugation in guanidine hydrochloride, urea, and lithium chloride. The subunits of the RNA were characterized by denaturing and non-denaturing electrophoresis in agarose gels. Poly(A)+-RNA, purified by oligo(dT)-cellulose affinity chromatography, was translated in a rabbit reticulocyte lysate assay and the products were immunoprecipitated with an experimentally infected mouse serum and a naturally infected human serum. After sodium dodecyl sulphate-polyacrylamide gel electrophoresis, fluorography of the polypeptides confirmed that the mRNA translated specific parasite antigens. |
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