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Over the past 30 years, the baculovirus-insect cell expression system has become one of the most widely used systems for routine production of recombinant proteins. Creative Enzymes actively conducts the evolvement of the viral vector system and the development of engineered insect cell lines. We provide professional services for enzyme expression in baculovirus-insect system. Our outstanding skills in vector design and cell engineering will significantly enhance the efficiency of your research with target enzymes.
When expressing cytosolic enzymes, membrane-bound proteins or enzymes with more complex post-translational modifications, baculovirus-insect cell expression system is preferably considered. Many technological improvements increase the popularity of this system, including the development of a wide variety of transfer vectors, simplified recombinant virus isolation and quantification methods, advances in cell culture technology, and the commercial availability of reagents. However, the advantages of the system are the key points taking into account.
In baculovirus-insect cell expression system, the recombinant enzymes can form proper eukaryotic posttranslational modifications without complication, including phosphorylation, N- and O-glycosylation, correct signal peptide cleavage, proper proteolytic processing, acylation, palmitylation, myristylation, amidation, carboxymethylation, and prenylation. As a result of these modifications, biologically and antigenically active molecules can be properly produced. Besides, unlike prokaryotic systems, correct protein folding and the formation of S-S bond can be achieved, which may become an obstacle for some large-sized enzyme expression.
The gene to be cloned is placed under the strong control of the viral polyhedrin promoter, allowing expression of heterologous enzyme of up to 30% of cell protein. Furthermore, combined with the optimal large-scale cultivation, the baculovirus insect cell system has potential to produce large quantities of recombinant enzymes.
The expression vectors are prepared from the baculovirus which can attack invertebrates but not vertebrates or plants, thus ensuring safety. The inherent safety for vertebrates makes baculovirus-insect system an attractive alternative to other higher eukaryotic expression system.
Figure 1: Baculovirus insect cell expression system. In cells infected with a wild-type virus (a) large amounts of polyhedra and p10 are formed. In most cases in the recombinant virus (b) the polyhedrin and/or p10 promoter is used to drive the expression of a foreign gene and the gene cassette replaces the polyhedrin gene, resulting in a recombinant with a polyhedron-negative phenotype and (c) high levels of recombinant protein are produced instead of polyhedrin.
Owing to these advantages, insect cells have been shown to produce near-authentic eukaryotic proteins and enzymes. Therefore, this expression system is widely used in biomedical research and in both medical and veterinary practice. One important application is the production of subunit vaccines based on baculovirus-expressed protein. Other applications are found in the expression of newly-developed enzymes to determine enzymatic properties.
With the professional technical support and the extensive experiences in molecular biology and fermentation, Creative Enzymes has shown the expertise in expressing enzymes in baculovirus-insect. According to your particular demands, Creative Enzymes offers expression and production in different scales. Each requested enzyme is produced under standard control with analysis certificates attached. Our expression service is guaranteed with both reliability and efficiency. Please contact us for any question about the service.
