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Schistosoma Mansoni Tropomyosin: Production and Purification of the Recombinant Protein and Studies on its Immunodiagnostic Potential

Han XuDepartments of Microbiology, Pathology, and Biochemistry, School of Medicine, State University of New York, Department of Epidemiology, School of Public Health, The University of Michigan, Buffalo, New York

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David M. RekoshDepartments of Microbiology, Pathology, and Biochemistry, School of Medicine, State University of New York, Department of Epidemiology, School of Public Health, The University of Michigan, Buffalo, New York

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William AndrewsDepartments of Microbiology, Pathology, and Biochemistry, School of Medicine, State University of New York, Department of Epidemiology, School of Public Health, The University of Michigan, Buffalo, New York

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Gene I. HigashiDepartments of Microbiology, Pathology, and Biochemistry, School of Medicine, State University of New York, Department of Epidemiology, School of Public Health, The University of Michigan, Buffalo, New York

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Laura NicholsonDepartments of Microbiology, Pathology, and Biochemistry, School of Medicine, State University of New York, Department of Epidemiology, School of Public Health, The University of Michigan, Buffalo, New York

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Philip T. LoverdeDepartments of Microbiology, Pathology, and Biochemistry, School of Medicine, State University of New York, Department of Epidemiology, School of Public Health, The University of Michigan, Buffalo, New York

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A cDNA that encodes Schistosoma mansoni tropomyosin, except for 10 amino acids at the amino terminus, has been cloned into a pOTSNCO plasmid vector. Induced expression resulted in a constant level of recombinant protein production. The recombinant S. mansoni tropomyosin was purified from preparative SDS-PAGE gel and by a combination of 20% ammonium sulfate fractionation and fast protein liquid chromatographyionexchange chromatography. The purified recombinant S. mansoni tropomyosin was tested as an immunodiagnostic reagent in Western blot and enzyme-linked immunosorbent assays. Sera from individual patients with chronic S. mansoni infection, but not S. haematobium, S. japonicum, parasitic infections other than schistosomiasis, and without infection reacted with the recombinant tropomyosin. The species specificity of S. mansoni tropomyosin suggests that further study of its potential as an immunodiagnostic reagent is warranted.

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