Expression and characterization of bovine lactoperoxidase by recombinant vaccinia virus |
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Authors: | Tetsuya Tanaka Xuenan Xuan Asato Kojima Ikuo Igarashi Kozo Fujisaki Kei-ichi Shimazaki |
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Institution: | (1) Laboratory of Emerging Infectious Diseases, Department of Frontier Veterinary Science, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan;(2) National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-855, Japan;(3) Department of Pathology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan;(4) Laboratory of Dairy Food Science, Research Faculty of Agriculture, Hokkaido University, Sapporo, Hokkaido 060-8589, Japan |
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Abstract: | Lactoperoxidase (LPO) is a 78 kDa heme-containing oxidation–reduction enzyme present in milk, found in physiological fluids
of mammals. LPO has an antimicrobial activity, and presumably contribute to the protective functions of milk against infectious
diseases. In this study, recombinant vaccinia virus expressing bovine LPO (vv/bLPO) was constructed. In rabbit kidney (RK13)
cells infected with vv/bLPO, recombinant bLPO was detected in both cell extracts and culture supernatants. Tunicamycin treatment
decreased the molecular weight of recombinant bLPO, indicating that recombinant bLPO contains a N-linked glycosylation site.
The replication of recombinant vaccinia viruses expressing bovine lactoferrin (vv/bLF) at a multiplicity of infection (moi)
of 5 plaque-forming units (PFU)/cell was inhibited by antiviral activity of recombinant bLF, suggesting that vv/bLF has an
antiviral effect against vaccinia virus. On the other hand, the replication of vv/bLPO at a moi of 5 PFU/cell was not inhibited
by antiviral activity of recombinant bLPO, indicating that this recombinant virus could be used as a suitable viral vector.
These results indicate that a combination of bLPO and vaccinia virus vector may be useful for medical and veterinary applications
in vivo. |
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Keywords: | Lactoperoxidase Lactoferrin N-linked glycosylation Vaccinia virus Antiviral effects |
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