title: Expression of recombinant human interferon-γ with antiviral activity in the bi-cistronic baculovirus-insect/larval system.
abstract: A bi-cistronic baculovirus-insect/larval system containing a polyhedron promoter, an internal ribosome entry site (IRES), and an egfp gene was developed as a cost-effective platform for the production of recombinant human interferon gamma (rhIFN-γ). There was no significant difference between the amounts of rhIFN-γ produced in the baculovirus-infected Spodoptera frugiferda 21 cells grown in serum-free medium and the serum-supplemented medium, while the Trichoplusia ni (T. ni) and Spodoptera exigua (S. exigua) larvae afforded rhIFN-γ amounting to 1.08±0.04 and 9.74±0.35 µg/mg protein respectively. The presence of non-glycosylated and glycosylated rhIFN-γ was confirmed by immunoblot and lectin blot. The immunological activity of purified rhIFN-γ, with 96% purity by Nickel (II)-nitrilotriacetic acid (Ni-NTA) affinity chromatography, was similar to that commercially available. Moreover, the rhIFN-γ protein from T. ni had more potent antiviral activity. These findings suggest that this IRES-based expression system is a simple and inexpensive alternative for large-scale protein production in anti-viral research.
authors: Institute of Molecular Medicine, National Tsing Hua University, Hsinchu, Taiwan, ROC.
authors: Wen-Shuo
authors: WS
authors: Chen
authors: Oliver B
authors: OB
authors: Villaflores
authors: Tzyy-Rong
authors: TR
authors: Jinn
authors: Ming-Tsair
authors: MT
authors: Chan
authors: Yen-Chung
authors: YC
authors: Chang
authors: Tzong-Yuan
authors: TY
authors: Wu
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D002846:Chromatography, Affinity; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000821:virology; D014176:Protein Biosynthesis; D012270:Ribosomes / Q000235:genetics*; D018411:Spodoptera / Q000235:genetics / Q000378:metabolism* / Q000821:virology*
chemical_list: C046697:glycosylated gamma interferon; D007371:Interferon-gamma