High level expression of biologically active canine interferon-alpha subtype 4 using a baculovirus

• Published in: Journal of Veterinary Medical Science Vol. 68; no. 12; pp. 1347 - 1349
• Main Authors: Ruttanapumma, R.(Kitasato Univ., Towada, Aomori (Japan). School of Veterinary Medicine and Animal Sciences), Anzai, M, Takegawa, M, Okamura, M, Maehara, N, Sato, K, Yoshioka, K, Itoh, A, Nakamura, M, Takehara, K
• Format: Journal Article
• Language: English
• Published: Japan JAPANESE SOCIETY OF VETERINARY SCIENCE 01.12.2006
• Subjects: Animals; ANTIMICROBIAL PROPERTIES; antiviral activity; BACULOVIRIDAE; Baculovirus; canine interferon-α; Cell Line; CHIEN; DOGS; expression; Gene Expression Regulation; Insecta; INTERFERON; Interferon-alpha - metabolism; INTERFERONAS; INTERFERONS; PERRO; PROPIEDADES ANTIMICROBIANAS; PROPRIETE ANTIMICROBIENNE; PURIFICACION; PURIFICATION; Vesicular stomatitis virus
• ISSN: 0916-7250; 1347-7439
• DOI: 10.1292/jvms.68.1347

In this study, a high amount of bioactive recombinant canine interferon-alpha subtype 4 (CaIFN-alpha4) was expressed in a baculovirus system. For easy purification, it was expressed as a CaIFN-alpha4 bearing histidine hexamer at the C-terminal region, designated CaIFN-alpha4His. CaIFN-alpha4His was detected in culture supernatants of insect cells infected with the recombinant virus using sodium dodecyl sulfate-polyarcylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue staining. The level of expression was very high, and approximately 1 mg of purified protein, with 5.0 x 10E7 units/mg, was obtained from 300 ml of culture supernatant. The purified product showed antiviral activity against vesicular stomatitis virus on canine tumor cell line A72 and chicken embryo fibroblast cells.