Expression of recombinant human interferon-γ with antiviral activity in the bi-cistronic baculovirus-insect/larval system
| Autor: | Wen-Shuo Chen, Tzong-Yuan Wu, Oliver B. Villaflores, Tzyy-Rong Jinn, Yen-Chung Chang, Ming-Tsair Chan |
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| Rok vydání: | 2011 |
| Předmět: |
Genetic Vectors
Gene Expression Spodoptera Applied Microbiology and Biotechnology Biochemistry Chromatography Affinity Analytical Chemistry law.invention Cell Line Interferon-gamma Affinity chromatography law Exigua medicine Trichoplusia Animals Humans Interferon gamma Molecular Biology biology fungi Organic Chemistry Gene Transfer Techniques General Medicine biology.organism_classification Molecular biology Blot Internal ribosome entry site Larva Protein Biosynthesis Recombinant DNA Baculoviridae Ribosomes Biotechnology medicine.drug |
| Zdroj: | Bioscience, biotechnology, and biochemistry. 75(7) |
| ISSN: | 1347-6947 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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