Advances in Insect Virology

King, Linda A.; Possee, Robert D.; Hughes, David S.; Atkinson, Allan E.; Palmer, Christopher P.; Marlow, Susan; Pickering, J.; Joyce, Kirsti A.; Lawrie, Alison M.; Miller, Davin; Beadle, David J.

doi:10.1016/s0065-2806(08)60062-4

Cited by 7 publications

(5 citation statements)

References 324 publications

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“…These viruses contain circular double-stranded DNA genomes of 90-160 kb in length [1][2][3]. Since its introduction in 1983 [4], the BEVS (baculovirus expression vector system) has become one of the most popular protein expression systems used in industry and molecular biology laboratories.…”

Section: Introductionmentioning

confidence: 99%

Enhancement of correct protein folding in vivo by a non-lytic baculovirus

Lee

et al. 2004

Biochemical Journal

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The BEVS (baculovirus expression vector system) is widely used for the production of proteins. However, engineered proteins frequently experience the problem of degradation, possibly due to the lytic nature of the conventional BEVS (herein referred to as L-BEVS). In the present study, a non-lytic BEVS (N-BEVS) was established by random mutagenesis of viral genomes. At 5 days post-infection, N-BEVS showed only 7 % cell lysis, whereas L-BEVS showed 60 % lysis of cells. The quality of protein expressed in both N-and L-BEVSs was examined further using a novel FRET (fluorescence resonance energy transfer)-based assay. To achieve this, we constructed a concatenated fusion protein comprising LUC (luciferase) sandwiched between EYFP (enhanced yellow fluorescent protein) and ECFP (enhanced cyan fluorescent protein). The distance separating the two fluorescent proteins in the fusion protein EYFP-LUC-ECFP (designated hereafter as the YLC construct) governs energy transfer between EYFP and ECFP. FRET efficiency thus reflects the compactness of LUC, indicating its folding status. We found more efficient FRET in N-BEVS compared with that obtained in L-BEVS, suggesting that more tightly folded LUC was produced in N-BEVS. YLC expression was also analysed by Western blotting, revealing significantly less protein degradation in N-BEVS than in L-BEVS, in which extensive degradation was observed. This FRETbased in vivo folding technology showed that YLC produced in N-BEVS is more compact, correlating with improved resistance to degradation. N-BEVS is thus a convenient alternative for L-BEVS for the production of proteins vulnerable to degradation using baculoviruses.

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Section: Introductionmentioning

confidence: 99%

Enhancement of correct protein folding in vivo by a non-lytic baculovirus

Lee

et al. 2004

Biochemical Journal

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“…In addition, we found evidence for four natural hosts ( D. pseudoobscura, D. subobscura, D. americana , D. cf. ananassae ) which have neither been reported as natural nor artificial hosts before [8] – [10] . Our results thus suggest that DCV is naturally infecting a much broader range of Drosophila host species than previously thought.…”

Section: Resultsmentioning

confidence: 99%

“…In contrast to the closely related Cricket paralysis virus (CrPV), which is highly similar to DCV in terms of viral morphology, genome size, and gene arrangement and which infects hosts in several insect orders [3] , [4] , the known host range of DCV is much more narrow [2] . DCV naturally infects D. melanogaster [5] , [6] and D. simulans [3] , [7] from the melanogaster subgroup, but natural infections in other species are unknown to date [8] – [10] . To test for susceptibility to DCV, Jousset [11] artificially infected 15 Drosophila species, four other dipterans and two lepidopterans by introducing DCV into the abdominal cavity.…”

Section: Introductionmentioning

confidence: 99%

Host Range and Specificity of the Drosophila C Virus

Kapun¹,

Nolte²,

Flatt³

et al. 2010

PLoS ONE

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BackgroundThe Drosophila C virus (DCV) is a common and well-studied Drosophila pathogen. Although natural infections are known from Drosophila melanogaster and D. simulans, and artificial infections have been reported from several Drosophila species and other insects, it remains unclear to date whether DCV infections also occur naturally in other Drosophila species.Methods/Principal FindingsUsing reverse transcription PCR, we detected natural infections in six Drosophila species, which have not been previously known as natural hosts. By subsequent Sanger sequencing we compared DCV haplotypes among eight Drosophila host species. Our data suggest that cross-infections might be frequent both within and among species within the laboratory environment. Moreover, we find that some lines exhibit multiple infections with distinct DCV haplotypes.ConclusionsOur results suggest that the natural host range of DCV is much broader than previously assumed and that cross-infections might be a common phenomenon in the laboratory, even among different Drosophila hosts.

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“…Baculoviruses are a diverse group of common insect pathogens that primarily infect the order Lepidoptera. These viruses contain circular double-stranded DNA genomes of 90 to 160 kb (6,30,38,41). They infect insect cells productively and generate numerous viral progenies.…”

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confidence: 99%

“…Persistent viral infection generates sporadic outbreaks in natural populations of infected insects that appear to be viral reactivation (17,33) caused by stress factors (26,30,33,42) such as overcrowding, lack of food (43), or thermal shock (24). The ingestion of heterologous viruses has also been shown to activate persistent viral infection (26,30,37,42). Despite these reports of occasional observations of persistently infected insects, the mechanisms that cause persistent baculovirus infection remain unknown.…”

mentioning

confidence: 99%

Persistent Baculovirus Infection Results from Deletion of the Apoptotic Suppressor Gene p35

1998

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Infection with the wild-type baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) results in complete death of Spodoptera frugiperda (Sf) cells. However, infection of Sf cells with AcMNPV carrying a mutation or deletion of the apoptotic suppressor gene p35 allowed the cloning of surviving Sf cells that harbored persistent viral genomes. Persistent infection established with the virus with p35 mutated or deleted was blocked by stable transfection of p35 in the host genome or by insertion of the inhibitor of apoptosis (iap) gene into the viral genome. These artificially established persistently virus-infected cells became resistant to subsequent viral challenge, and some of the cell lines carried large quantities of viral DNA capable of early gene expression. Continuous release of viral progenies was evident in some of the persistently virus-infected cells, and transfection of p35 further stimulated viral activation of the persistent cells, including the reactivation of viruses in those cell lines without original continuous virus release. These results have demonstrated the successful establishment of persistent baculovirus infections under laboratory conditions and that their establishment may provide a novel continuous, nonlytic baculovirus expression system in the future.

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Advances in Insect Virology

Cited by 7 publications

References 324 publications

Enhancement of correct protein folding in vivo by a non-lytic baculovirus

Enhancement of correct protein folding in vivo by a non-lytic baculovirus

Host Range and Specificity of the Drosophila C Virus

Persistent Baculovirus Infection Results from Deletion of the Apoptotic Suppressor Gene p35

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