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Identification of Epitope on DNA-binding Protein Expressed in Insect Cell Infected by Baculovirus

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Abstract

DNA-binding protein (DBP) is an early gene product produced during viral replication. Polyclonal anti-DBP was produced using rabbit by intradermal injections of Escherichia coli-expressed purified recombinant DBP. Prepared anti-DBP completely blocked the replication of baculovirus in insect cells. The anti-DBP binding to DBP was confirmed by both Western blotting with Tn-5B1-4 insect cell lysates as well as immunostained baculovirus-infected Tn-5B1-4 insect cells. To determine the anti-DBP epitope 12 peptides were synthesized and their specific-binding activities were measured using ELISA. Based on specific-binding activity against anti-DBP the epitope was predicted to be between amino acid residues 248–265 (QRMSVEDFDRLFEMDKID). Especially from 18 amino acid residues it was further to be narrowed between amino acid residues 260–265 (EMDKID) which showed a critical role in specific-binding activity.

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Authors and Affiliations

  1. Laboratory of Biotechnology, Shizuoka University, 836 Ohya, Shizuoka, 422-8529, Japan

    Mami Suzuki & Enoch Y. Park

  2. Laboratory of Animal Reproduction and Physiology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka, 422-8529, Japan

    Tetsuya Kohsaka

Authors
  1. Mami Suzuki
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  2. Tetsuya Kohsaka
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  3. Enoch Y. Park
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Correspondence to Enoch Y. Park.

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Suzuki, M., Kohsaka, T. & Park, E.Y. Identification of Epitope on DNA-binding Protein Expressed in Insect Cell Infected by Baculovirus. Mol Biol Rep 33, 97–102 (2006). https://doi.org/10.1007/s11033-005-0414-0

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  • DOI: https://doi.org/10.1007/s11033-005-0414-0

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