Baculovirus-mediated protein expression is a robust experimental technique for producing recombinant higher-eukaryotic proteins because it combines high yields with considerable post-translational modification capabilities. In this expression system, the determination of the titer of recombinant baculovirus stocks is important to achieve the correct multiplicity of infection for effective ampli...

Baculovirus vectors can efficiently transduce human hepatoma cells and primary hepatocytes in culture. We report the potential use of baculovirus as a vector for gene delivery into cells of the mammalian central nervous system. We generated a “pseudotyped”baculovirus encoding the bacterialβ-galactosidase(β-Gal)gene(LacZ),under the control of the cytomegalovirus promoter, and the vesicular stoma...

The 579-nucleotide 5' untranslated region (5'UTR) of the Rhopalosiphum padi virus (RhPV) possesses a cross-kingdom internal ribosome entry site (IRES) activity that functions in insect, mammalian, and plant-derived in vitro translation systems, and six TAAG motifs within the DNA fragment encoding the RhPV 5'UTR were previously found to confer the RhPV 5'UTR with late promoter activity in baculo...

Baculoviruses are arthropod‐specific, enveloped viruses with circular, supercoiled double‐ stranded DNA genomes [1]. They infect Lepidoptera (butterflies and moths), Hymenoptera (sawflies) and Diptera (mosquitoes) [2]. While many viruses are studied because of their damaging effects, the study of baculoviruses was stimulated by their potential utility to control insect pests [3]. Later, the uti...

Recombinant protein expression using the Baculovirus Expression System is widely used for the production of therapeutics, diagnostics, and reagents. The current “gold standard” for Baculovirus titration, the plaque assay, is labor intensive and time consuming. The realtime PCR assay has the advantage of speed and accuracy. We determined Baculovirus tire by a novel quantitative PCR (qPCR) method...

Detailed investigation of virus replication is facilitated by the construction of a full-length infectious clone of the viral genome. To date, this has not been achieved for members of the family Dicistroviridae. Here we demonstrate the construction of a baculovirus that expresses a dicistrovirus that is infectious in its natural host. We inserted a full-length cDNA clone of the genomic RNA of ...

(2)-b-2*,3*-Dideoxy-3*-thiacytidine (lamivudine [3TC]) is a nucleoside analog which effectively interferes with the replication of hepatitis B virus (HBV) DNA in vitro and in vivo. We have investigated the antiviral properties of 3TC in vitro in HepG2 cells infected with recombinant HBV baculovirus. Different types of information can be obtained with the HBV baculovirus-HepG2 system because (i)...

Recombinant baculoviruses have established themselves as a favoured technology for the high-level expression of recombinant proteins. The construction of recombinant viruses, however, is a time consuming step that restricts consideration of the technology for high throughput developments. Here we use a targeted gene knockout technology to inactivate an essential viral gene that lies adjacent to...

The baculovirus expression vector system has been commonly employed as a rapid and inexpensive approach for recombinant protein production in insect cells (1, 2). Despite the wide applications, baculovirus infection results in insect cell death and lysis in a few days post-infection, which can lead to suboptimal protein processing late in the infection phase. In addition, some proteins that are...

The Baculoviridae is a diverse family of pathogens that are infectious for arthropods and are characterized by a complex replication cycle that culminates in the occlusion of virions in a crystalline protein matrix (Blissard & Rohrmann, 1990). Over 400 lepidopteran species serve as hosts for these viruses, with a single virus isolate usually restricted to one or a few related species. In additi...