Production of Human Beta Interferon in Insect Cells Infected with a Baculovirus Expression Vector

Abstract

Autographa californica nuclear polyhedrosis virus (AcNPV) was used as an expression vector for human beta interferon. By using specially constructed plasmids, the protein-coding sequences for interferon were linked to the AcNPV promoter for the gene encoding for polyhedrin, the major occlusion protein. The interferon gene was inserted at various locations relative to the AcNPV polyhedrin transcriptional and translational signals, and the interferon-polyhedrin hybrid genes were transferred to infectious AcNPV expression vectors. Biologically active interferon was produced, and greater than 95% was secreted from infected insect cells. A maximum of ca. 5 X 10(6) U of interferon activity was produced by 10(6) infected cells. These results demonstrate that AcNPV should be suitable for use as a eucaryotic expression vector for the production of products from cloned genes.

Solutions against emerging infectious and noninfectious human diseases through the application of baculovirus technologies

“…Therefore, the versatility manifested by AcMNPV in tolerating genomic changes and the possibilities for scaling-up in cell culture not only enabled the use of this baculovirus as a biopesticide (Vail et al 1971), but also favored the development of innovations for the expression of recombinant proteins (Smith et al 1992) and VLPs (Pearson and Polly 1993) for the generation of protein-display platforms (Boublik et al 1995), or for use as a virus vector in gene deliveries to mammalian cells (Hofmann et al 1995). Accordingly, BEVS and BacMam technologies were born in just over 10 years between 1983 and 1995, and during that time, everything was prepared for their transfer to the human-and veterinary-health industries.…”

Structure Determination of Microtubules and Pili: Past, Present, and Future Directions

“…With the advent of single crystal X-ray diffraction (Kendrew et al, 1958), from the 1960s structural studies of soluble proteins became the principal focus due to the delivery of atomic/subatomic resolutions. In the 1980s the first soluble protein was determined by solution state nuclear magnetic resonance (NMR) spectroscopy (Williamson et al, 1985) and with the development of recombinant protein expression systems (Itakura et al, 1977;Smith et al, 1983;Cregg et al, 1993), by the mid-1990s there became a clear exponential rise in soluble/globular macromolecular structures being deposited in the protein data bank (Berman et al, 2000). Likewise, new methods for isolating and reconstituting membrane proteins has led to significant numbers of these structures (Vinothkumar and Henderson, 2010), primarily elucidated by crystallographic methods, being deposited in the PDB since the 2000s.…”

Improvement of protein production by engineering a novel antiapoptotic baculovirus vector to suppress the expression ofSf‐caspase‐1andTn‐caspase‐1

“…Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the most widely used baculovirus species (Oers, 2011), which has wide applications in virological research and biotechnology. Since the first successful utilization of baculovirus expression vector system (BEVS) in the expression of human β‐interferon in 1983 (Smith et al, 1983), a large number of recombinant proteins have been produced in baculovirus‐infected insect cells. As a powerful eukaryotic expression system, BEVS possesses many advantages, such as high expression level of recombinant proteins, proper posttranslational modifications for correct folding and specific biological function, high safety as it is non‐pathogenic to mammalian animals, and lower cost than mammalian cell expression systems that is suitable for large‐scale industrial production.…”