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Expression and characterization of the multiplied, recombinant preS1 antigen of hepatitis B virus

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Summary

The amino acid sequence encoded by the preS1 region of hepatitis B virus genome is expressed on the surface of virions and subviral particles. The preS1 region is involved in the recognition of specific receptors responsible for the attachment of HBV to the host cell. The cell receptor binding site was assigned to the preS1 (20–47 aa) fragment. In order to obtain a large quantity of preS1 binding domains of HBV the expression vector pWX4 was constructed. It contains four tandemly joined DNA sequences, each coding for preS1 (20–49 aa), fused with the 3′ end of a DNA fragment coding for 450 aa of β-galactosidase.E. coli cells transformed with this vector produce fusion protein β-gal-preSlx4 in the form of inclusion bodies. Owing to the specific trypsin digestion, the preSlx4 domain was cleaved from the fusion protein. The resulting product, a 16 kDa protein, was isolated and purified by anion exchange chromatography. The presence of four Asp-Pro bonds in this sequence and the primary structure of the first 28 N-terminal amino acids were determined. Following the confirmation of the antigenic properties, the recombinant preS1 protein was used for detection of the anti-preS1 response in sera from HBV infected patients.

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Authors and Affiliations

  1. Department of Biochemistry, Medical University of Łódz, Poland

    M. Sidorkiewicz

  2. Laboratory of Genetic Engineering, Sieradz

    G. Płucienniczak

  3. Polish Academy of Sciences, Center of Microbiology and Virology, Łódz, Poland

    A. Płucienniczak

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  1. M. Sidorkiewicz
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  2. G. Płucienniczak
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  3. A. Płucienniczak
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Sidorkiewicz, M., Płucienniczak, G. & Płucienniczak, A. Expression and characterization of the multiplied, recombinant preS1 antigen of hepatitis B virus. Archives of Virology 140, 1935–1944 (1995). https://doi.org/10.1007/BF01322683

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  • DOI: https://doi.org/10.1007/BF01322683

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