High level expression of biologically active canine interferon-alpha subtype 4 using a baculovirus.
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 10(7) 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.
Ruttanapumma
Ruttapong
R
Laboratory of Zoonoses, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori, Japan.
Anzai
Manami
M
Takegawa
Masaoki
M
Okamura
Masashi
M
Maehara
Nobutoshi
N
Sato
Ko
K
Yoshioka
Kuniaki
K
Itoh
Akira
A
Nakamura
Masayuki
M
Takehara
Kazuaki
K
D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D004285:Dogs; D005786:Gene Expression Regulation; D007313:Insecta; D016898:Interferon-alpha / Q000378:metabolism*
D016898:Interferon-alpha