High level expression, purification, and in vivo activity of bovine granulocyte-colony stimulating factor produced using a baculovirus system.
A bovine granulocyte-colony stimulating factor (bG-CSF) cDNA clone bearing a C-terminal poly-His-tag (bG-CSFHis) was constructed and expressed by the baculovirus expression system. The bG-CSFHis was expressed as an approximately 19kDa protein in the culture supernatants and was purified using a nickel chelate column. The purified bG-CSFHis had bioactivity in vitro in the NFS-60 bioassay. In order to evaluate activity in vivo, purified bG-CSFHis was administered to cattle as single or multiple dosages. The bG-CSFHis increased neutrophil counts in peripheral blood and modulated the phagocytic activity of the neutrophils. The data indicates that the recombinant protein had activity in vivo.
Nagata
Tomoshi
T
Laboratory of Poultry Diseases, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, Aomori 034-8628, Japan.
Tohda
Yukihiko
Y
Yokomizo
Yuichi
Y
Nakamura
Masayuki
M
Takehara
Kazuaki
K
D000818:Animals; D016367:Baculoviridae / Q000737:chemistry / Q000235:genetics; D001681:Biological Assay; D002417:Cattle / Q000276:immunology*; D045744:Cell Line, Tumor / Q000276:immunology; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007958:Leukocyte Count / Q000662:veterinary; D051379:Mice; D010587:Phagocytosis / Q000276:immunology; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D018411:Spodoptera / Q000235:genetics
D018076:DNA, Complementary; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D012313:RNA