Characterization of a new Autographa californica multiple nucleopolyhedrovirus (AcMNPV) polyhedra mutant

Ribeiro, Bergmann Morais; Generino, Ana Paula Montenegro; Acacio, Cláudia Natércia Lima; Kalapothakis, Evanguedes; Báo, Sônia Nair

doi:10.1016/j.virusres.2008.10.010

Cited by 11 publications

(2 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We observed only one band when crystals formed by Mola-Granulin were resolved ( Figure 6 E, lane 2) and several faint bands together with a strongest band for crystals formed by AcMNPV-Polyhedrin ( Figure 6 E, lane 4). The crystal structure of baculoviruses’ OBs has been shown to be dependent of the amino acid sequence of the main occlusion body protein [ 42 , 43 , 44 , 45 ] and also on interactions with other viral and/or host proteins [ 46 ].…”

Section: Resultsmentioning

confidence: 99%

A Novel Betabaculovirus Isolated from the Monocot Pest Mocis latipes (Lepidoptera: Noctuidae) and the Evolution of Multiple-Copy Genes

Ardisson-Araújo

Silva

Melo

et al. 2018

Viruses

View full text Add to dashboard Cite

In this report, we described the genome of a novel baculovirus isolated from the monocot insect pest Mocis latipes, the striped grass looper. The genome has 134,272 bp in length with a G + C content of 38.3%. Based on the concatenated sequence of the 38 baculovirus core genes, we found that the virus is a betabaculovirus closely related to the noctuid-infecting betabaculoviruses including Pseudaletia unipuncta granulovirus (PsunGV), Trichoplusia ni granulovirus (TnGV), Helicoverpa armigera granulovirus (HearGV), and Xestia c-nigrum granulovirus (XecnGV). The virus may constitute a new Betabaculovirus species tentatively named Mocis latipes granulovirus (MolaGV). After gene content analysis, five open reading frames (ORFs) were found to be unique to MolaGV and several auxiliary genes were found including iap-3, iap-5, bro-a, bro-b, and three enhancins. The virus genome lacked both chitinase and cathepsin. We then looked at the evolutionary history of the enhancin gene and found that betabaculovirus acquired this gene from an alphabaculovirus followed by several duplication events. Gene duplication also happened to an endonuclease-like gene. Genomic and gene content analyses revealed both a strict collinearity and gene expansion into the genome of the MolaGV-related species. We also characterized the granulin gene using a recombinant Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and found that occlusion bodies were produced into the nucleus of infected cells and presented a polyhedral shape and no occluded virions within. Overall, betabaculovirus genome sequencing is of importance to the field as few genomes are publicly accessible. Mocis latipes is a secondary pest of maize, rice, and wheat crops in Brazil. Certainly, both the discovery and description of novel baculoviruses may lead to development of greener and safer pesticides in order to counteract and effectively control crop damage-causing insect populations

show abstract

Section: Resultsmentioning

confidence: 99%

A Novel Betabaculovirus Isolated from the Monocot Pest Mocis latipes (Lepidoptera: Noctuidae) and the Evolution of Multiple-Copy Genes

Ardisson-Araújo

Silva

Melo

et al. 2018

Viruses

View full text Add to dashboard Cite

show abstract

“…For immunoblotting, samples were resolved by 12% SDS-PAGE and transferred onto a nitrocellulose membrane (Sigma) using the Trans-Blot® SD – Semi Dry Transfer Cell (BioRad). The membrane was then blocked in 1× PBS Buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 2 mM KH 2 PO 4 , pH 7.4) containing 3% skimmed milk powder for 16 h at 4°C, washed three times with PBS tween (0.05%) and probed with (i) mouse monoclonal anti-hexa-histidine (anti-6xHis) antibody (Sigma), (ii) rabbit anti-Polh antiserum [31], or (iii) anti-Polh-GarMbFV-CP-6xHis (produced in this work) followed by incubation with the alkaline phosphatase-conjugated anti-mouse/rat or anti-rabbit secondary antibodies (Sigma). Blots were developed using the NBT/BCIP (Sigma) substrate dissolved in alkaline phosphatase buffer (NaCl 100 mM, MgCl 2 5 mM e Tri-HCl 100 mM – pH 9.0).…”

Section: Methodsmentioning

confidence: 99%

A baculovirus-mediated strategy for full-length plant virus coat protein expression and purification

Ardisson-Araújo

Rocha

Hedil

et al. 2013

Virol J

Self Cite

View full text Add to dashboard Cite

BackgroundGarlic production is severely affected by virus infection, causing a decrease in productivity and quality. There are no virus-free cultivars and garlic-infecting viruses are difficult to purify, which make specific antibody production very laborious. Since high quality antisera against plant viruses are important tools for serological detection, we have developed a method to express and purify full-length plant virus coat proteins using baculovirus expression system and insects as bioreactors.ResultsIn this work, we have fused the full-length coat protein (cp) gene from the Garlic Mite-borne Filamentous Virus (GarMbFV) to the 3′-end of the Polyhedrin (polh) gene of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). The recombinant baculovirus was amplified in insect cell culture and the virus was used to infect Spodoptera frugiperda larvae. Thus, the recombinant fused protein was easily purified from insect cadavers using sucrose gradient centrifugation and analyzed by Western Blotting. Interestingly, amorphous crystals were produced in the cytoplasm of cells infected with the recombinant virus containing the chimeric-protein gene but not in cells infected with the wild type and recombinant virus containing the hexa histidine tagged Polh. Moreover, the chimeric protein was used to immunize rats and generate antibodies against the target protein. The antiserum produced was able to detect plants infected with GarMbFV, which had been initially confirmed by RT-PCR.ConclusionsThe expression of a plant virus full-length coat protein fused to the baculovirus Polyhedrin in recombinant baculovirus-infected insects was shown to produce high amounts of the recombinant protein which was easily purified and efficiently used to generate specific antibodies. Therefore, this strategy can potentially be used for the development of plant virus diagnostic kits for those viruses that are difficult to purify, are present in low titers or are present in mix infection in their plant hosts.

show abstract

Biotechnological applications of occlusion bodies of Baculoviruses

López

Diez

Alfonso

et al. 2018

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

The ability of Baculoviruses to hyper-express very late genes as polyhedrin, the major component of occlusion bodies (OBs) or polyhedra, has allowed the evolution of a system of great utility for biotechnology. The main function of polyhedra in nature is to protect Baculovirus in the environment. The possibility of incorporating foreign proteins into the crystal by fusing them to polyhedrin (POLH) opened novel potential biotechnological uses. In this review, we summarize different applications of Baculovirus chimeric OBs. Basically, the improvement of protein expression and purification with POLH as a fusion partner; the use of recombinant polyhedra as immunogens and antigens, and the incorporation of proteins into polyhedra to improve Baculoviruses as bioinsecticides. The results obtained in each area and the future trends in these topics are also discussed.

show abstract

Characterization of a new Autographa californica multiple nucleopolyhedrovirus (AcMNPV) polyhedra mutant

Cited by 11 publications

References 25 publications

A Novel Betabaculovirus Isolated from the Monocot Pest Mocis latipes (Lepidoptera: Noctuidae) and the Evolution of Multiple-Copy Genes

A Novel Betabaculovirus Isolated from the Monocot Pest Mocis latipes (Lepidoptera: Noctuidae) and the Evolution of Multiple-Copy Genes

A baculovirus-mediated strategy for full-length plant virus coat protein expression and purification

Biotechnological applications of occlusion bodies of Baculoviruses

Contact Info

Product

Resources

About