Expression of Recombinant Human Interferon-γ with Antiviral Activity in the Bi-Cistronic Baculovirus-Insect/Larval System

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概要

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• 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.

著者

• Jinn Tzyy-rong

  Graduate Institute Of Biotechnology National Chung Hsing University

• Wu Tzong-yuan

  Department Of Bioscience Technology Chung Yuan Christian University

• CHEN Wen-Shuo

  Institute of Molecular Medicine, National Tsing Hua University

• VILLAFLORES Oliver

  Department of Chemistry, Chung Yuan Christian University

• CHAN Ming-Tsair

  Institute of BioAgricultural Sciences, Academia Sinica

• CHANG Yen-Chung

  Institute of Molecular Medicine, National Tsing Hua University

• Villaflores Oliver

  Department Of Chemistry Chung Yuan Christian University

• Chang Yen-chung

  Institute Of Molecular Medicine National Tsing Hua University

• Jinn Tzyy-rong

  Graduate Institute Of Chinese Medical Science China Medical University

関連論文

• Functional Expression of FIP-gts, a Fungal Immunomodulatory Protein from Ganoderma Tsugae in Sf21 Insect Cells
• Expression and Immunological Studies of Classical Swine Fever Virus Glycoprotein E2 in the Bi-Cistronic Baculovirus/Larvae Expression System
• Expression of Recombinant Human Interferon-γ with Antiviral Activity in the Bi-Cistronic Baculovirus-Insect/Larval System
• Expression of Recombinant Human Interferon-γ with Antiviral Activity in the Bi-Cistronic Baculovirus-Insect/Larval System
• Functional Expression and Characterization of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis in Sf21 Insect Cells

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