Baculoviruses as vectors for foreign gene expression in insect cells

Abstract

Recombinant DNA technology has enabled the expression of a large number of eukaryotic genes in vitro. There has been growing interest in recent years in the development of baculovirus expression vectors as an easily manipulated gene expression system. Both virus and insect cell culture are relatively easy to handle, and biologically active proteins have been produced abundantly from a variety of eukaryotic, prokaryotic and viral genes. This review describes the molecular biology of baculoviruses and some of the current applications of the baculovirus expression system in insect cells. Specific emphasis is placed on those features relevant to the use of this system in pesticide research.

Synthesis of functional GABAA receptors in stable insect cell lines

“…St9.G~l cells (0.5 x 106) were placed in 60 mm Petri dishes and equilibrated in a bathing saline (120 mM NaCl, 4 mM KCl, 10 mM CaCl,, 11 mM MgC&, 10 mM HEPES, pH 7.2). Patch electrodes (3)(4)(5) were filled with 110 mM KCI, 1 mM CaCl,, 10 mM EGTA, 10 mM HEPES, pH 7.2. GABA (lo-' M; 1-2 s) was applied to the cells by pressure ejection (Picospritzer, General Valve Corporation) from 1 PM diameter micropipettes positioned 3&50prn from the whole-cell clamped cells.…”

Effects of Oxygen/Glucose/Glutamine Feeding on Insect Cell Baculovirus Protein Expression: A Study on Epoxide Hydrolase Production

“…More than 300 heterologous proteins have been expressed in the insect cell baculovirus expression system (Agathos, 1991;Atkinson et al, 1990;Luckow and Summers, 1988). This system has several advantages, including the overproduction of functional heterologous proteins owing to proper post-translational modifications and a strong polyhedrin promoter (Cameron et al, 1990;Fraser, 1989;Luckow and Summers, 19881, the ease of cell propagation (Agathos, 1991) due to the earlier development of 'mammalian cell cultures, and the minimization of environmental health and safety concern due to host-range limitations of the recombinant virus (Cameron et al, 1990;Fraser, 1989;Luckow and Summers, 1988).…”

Optimization of the Strategy for Recombinant Baculovirus Infection of Suspended Insect Cells

“…In vitro infection of insect cells with baculoviruses is increasingly considered as a viable means for the production of biopesticides, recombinant veterinary vaccines and other recombinant products (Atkinson et al, 1990). Over the last two decades, much effort has been focused on identifying factors that affect the productivities of such expression systems, including the cell line (Wickham et al, 1995), medium (Caron et al, 1990), oxygen supply (Gotoh et al, 2002), reactor design (Rice et al, 1993;Zhang et al, 1993) and infection strategy, which involves the multiplicity of infection (MOI), time of infection (TOI), initial cell density (ICD) and medium replacement (Carinhas et al, 2009;Hu and Bentley, 2001;Radford et al, 1997;Rodas et al, 2005;Yamaji et al, 1999;Zhang et al, 2005).…”