3. Gene Delivery and Expression Series Overexpression of Recombinant Protein in Large-scale Culture Using a Baculovirus-insect Cell Expression System: Based on the Study of Xanthine Oxidoreductase (3)
Division of Medical and Biological Chemistry, Graduate School of Medicine, Nippon Medical School
The baculovirus-insect cell system is widely used to express recombinant proteins. Posttranslational modifications of proteins, such as glycosylation and the phosphorylation and incorporation of cofactors, in Escherichia coli are less sufficient than those in mammalian cells. Unfortunately, mammalian cells are not appropriate for large-scale culture. However, insect cells are eukaryotic cells and are suitable for large-scale culture; they can be used to produce recombinant proteins that cannot be expressed in E. coli. We established a baculovirus-insect cell system to express xanthine oxidoreductase and obtained about ∼4 mg of purified enzyme from 2 to 4 L of culture medium. This amount of enzyme was sufficient for X-ray crystal structureanalysis. Here, we describe of the outline of our methods for culturing insect cells, producing recombinant baculoviruses, and expressing recombinant proteins in large-scale culture.
úãåãï 2012; 8(1), 26-30
insect cells, sf9, baculovirus, xanthine oxidoreductase
Ken Okamoto, Division of Metabolism and Nutrition, Department of Biochemistry and Molecular Biology, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku Tokyo 113-8602, Japan