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30 results for “Baculoviruses”
Fig. 5 in Successful transcription but not translation or assembly of Solenopsis invicta virus 3 in a baculovirus-driven expression system
Fig. 5. Confirmation of heterologous expression of SINV-3 transcript by amplification of the 3' (A) and 5' termini (B) from RNA templates purified from SINV- 3-transfected Sf21 cells. Regions amplified are illustrated in the genome diagram between (A) and (B). (A) Three plaque preparations (AcSINV-3 CiC, AcSINV-3 CiD, and AcSINV-3 DiD) were separated by centrifugation into soluble and pelleted fractions, treated with DNase I, reverse transcribed, and the 3' end of the genome amplified by PCR. Lane assignments were as follows: 1 = mass marker (bp); 2, 6 = AcSINV-3 CiC; 3, 7 = AcSINV-3 CiD; 4, 8 = AcSINV-3 DiD; 5, 9 = mock infection; 10 = positive control (wild-type virus); 11 = negative control; 12 = non-template control. (B) AcSINV-3 plaque preparations (CiC and DiD) evaluated by PCR of the 5' end of the genome (RNA preparations). Lane assignments were as follows: 1 = mass marker (bp); 2, 3 = DNase treated, reverse transcribed; 4, 5 = without DNase treatment, reverse transcribed; 6, 7 = DNase treated, without reverse transcription; 8, 9 = without DNase treatment, without reverse transcription; 10 = positive control; 11 = negative control; 12 = non-template control. (C) PCR amplification of the entire SINV-3 genome from DNA preparations of AcSINV-3 CiC (lane 2) and AcSINV-3 DiD (lane 3). Lane 1, molecular markers; lane 4, non-template control. (D) Western blot to evaluate translation of the SINV-3 transcript by detection of viral capsid protein 2 (VP2). Lane assignments were as follows: 1 = AcSINV-3 CiC (4 dpi); 2 = AcSINV-3 CiC (5 dpi); 3 = AcSINV-3 CiD (4 dpi); 4 = AcSINV-3 DiD (5 dpi); 5 = mock infection (negative control); 6 = positive control (purified wild-type SINV-3; kDa).
Fig. 3 in Successful transcription but not translation or assembly of Solenopsis invicta virus 3 in a baculovirus-driven expression system
Fig. 3. (A) Plaque assay results for recombinant SINV-3 (AcSINV-3) transfection of Sf21 cells indicating the dilution used for each plate. (B upper panel) Representa- tive plaque with Sf21 cells stained with neutral red 10 d afer transfection (magnified 100 times). (B lower panel) Corresponding mock-infected Sf21 cells (negative control) afer 10 d of exposure. Plaque areas identify infection of insect cells by virus with corresponding cell death.
Fig. 2 in Successful transcription but not translation or assembly of Solenopsis invicta virus 3 in a baculovirus-driven expression system
Fig. 2. (A) pFastBac1_SINV-3 hybrid construct map. Locations of the restriction sites (black hash marks), bacterial transposon Tn7 sites (grey triangles), polyhedrin promoter (angled arrow corresponding to the sequence below), SINV-3 open reading frames (dark closed arrows), and approximate location of the area detected by the polyclonal antibody preparation (pAb) are shown. (B) Verified sequence of the pFastBac1_SINV-3 hybrid construct illustrating the late gene polyhedrin promoter (angled arrow). SINV-3 sequence is in bold font,with pFastBac1 sequence in normal font, and restriction sites are superscripted and corresponding sequences italicized.Underlined sequence represents the inserted late gene polyhedrin core promoter.Analyzed sequences of the 5'and 3'termini and restriction sites were identical to the wild-type virus.
Fig. 1 in Successful transcription but not translation or assembly of Solenopsis invicta virus 3 in a baculovirus-driven expression system
Fig. 1. Schematic of the SINV-3 genome and sub-cloning strategy to assemble the pFastBac1 donor plasmid/SINV-3 construct. (A) Organization of the SINV-3 genome illustrating the 2 ORFs numbered 1 and 2 that encode for non-structural and structural proteins, respectively, and the genome sections sub-cloned. Oligonucleotide primers used to generate cDNA and amplify each section are indicated. Primers with introduced restriction sites are also indicated. (B) Unique restriction sites for each sub-clone and the assembly process employed to concatenate the entire SINV-3 genome in the pFastBac1 donor vector.
Full-length HTT purification from HEK293T suspension culture using baculovirus transduction protocol for over-expression 2018/04/02
<p>Structure-function open lab notebook project. Full-length HTT purification from HEK293T suspension culture using baculovirus transduction protocol for over-expression 2018/04/02.</p>
Fig. 1 in Genetic identification and biological characterization of Baculovirus isolated from Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil
Fig. 1. Phylogenetic relationships among HearNPV isolates (BR1, BR2, BR3, Br4, and GEM) based on analysis of concatenated nucleotide sequences of the lef-8 and lef-9. The evolutionary history was inferred by using the Maximum Likelihood method. The tree with the highest log likelihood (−799.3699) is shown. The percentage of trees in which the associated taxa clustered together is shown next to the branches. Initial tree(s) for the heuristic search were obtained by applying the Neighbor-Joining method to a matrix of pairwise dis- tances estimated using the Maximum Composite Likelihood (MCL) approach. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. The analysis involved 14 nucleotide sequences. All posi- tions containing gaps and missing data were eliminated.
Fig. 2 in Genetic identification and biological characterization of Baculovirus isolated from Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil
Fig. 2. Survival curves of Helicoverpa armigera (Lepidoptera: Noctuidae) afer feeding on corn leaves inoculated with 1 × 106 polyhedra per mL of baculovirus isolated. Gemstar®LC, BR1, BR2, BR3, BR4, and in control obtained with KaplanMeier estimator.
Large scale expression and purification of full length huntingtin Q46 from baculovirus expression system (2016/09/04)
<p>Open lab notebook huntingtin structure function project.<br> </p>
Test expression and purification of full length huntingtin Q23, Q46 and Q78 in baculovirus expression system (2016/08/04)
<p>Open lab notebook huntingtin structure function project.</p>
Synchronization of in vivo MacoNPV-A baculovirus infection by analysis of individual Mamestra configurata larval guts
<p><span>Many studies have examined the gene expression of baculoviruses during host infection by infecting cultured insect host cells; however, only a few have attempted to characterize the interaction between baculoviruses and insect larvae, which more accurately models the virus-insect relationship. The greatest challenge in assessing gene expression profiles <em>in vivo</em> in the larval gut is the lack of infection synchronization compared to cultured cells. Working with bertha armyworm, <em>Mamestra configurata</em>, larvae infected with the baculovirus Mamestra configurata nucleopolyhedrovirus-A<em> </em><span> </span>(MacoNPV-A), viral gene expression was measured using droplet-digital PCR showing that the rate of infection in individual insects varies extremely widely. Subsequent RNA-seq of individual guts revealed that gene expression patterns were consistent in individuals with similar levels of viral gene expression. Therefore, the best approach for analyzing baculovirus gene expression <em>in vivo</em> is to use the expression of an early viral gene as a means to select individuals that are closely matched in infection progress to produce the most synchronized infection cohorts for measuring gene expression profiles. In this way, a profile of gene expression was identified very early in infection that would have been masked by the activity of a few very robustly infected individuals present in sample pools made up of multiple individuals.</span></p>
Data from: Phenotypic variation in overwinter environmental transmission of a baculovirus and the cost of virulence
A pathogen's ability to persist in the environment is an ecologically important trait, and variation in this trait may promote coexistence of different pathogen strains. We asked whether naturally occurring isolates of the baculovirus that infects gypsy moth larvae varied in their overwinter environmental transmission, and whether this variation was consistent with a tradeoff or an upper limit to virulence that might promote pathogen diversity. We used experimental manipulations to replicate the natural overwinter infection process using 16 field-collected isolates. Virus isolates varied substantially in the fraction of larvae infected, leading to differences in overwinter transmission rates. Furthermore, isolates that killed more larvae also had higher rates of early larval death in which no infectious particles were produced, consistent with a cost of high virulence. Our results thus support the existence of a cost that could impose an upper limit to virulence even in a highly virulent pathogen.
Data from: Pathogen persistence in the environment and insect-baculovirus interactions: disease-density thresholds, epidemic burnout, and insect outbreaks
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Data from: Phenotypic variation in overwinter environmental transmission of a baculovirus and the cost of virulence
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Data from: Baculovirus infection triggers a positive phototactic response in caterpillars (a response to Dobson et al. Biol Letters 2015).
We recently reported that baculovirus Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) triggers positive phototaxis in Spodoptera exigua larvae, leading to death at elevated positions. Dobson et al. [1] (University of Stirling, Scotland) question our interpretation of the data. Unfortunately, Dobson et al. rely on unwarranted assumptions possibly reflecting a poor understanding of baculovirus–insect pathobiology, make invalid comparisons and fail to take relevant literature into account. Here, we recapitulate the context and interpretation of our experiments and highlight the misinterpretations by Dobson et al.
Large scale expression and purification of full length huntingtin Q23 from baculovirus expression system (2016/08/29)
<p>Open lab notebook huntingtin structure function project.<br> </p>
Fig. 4 in Successful transcription but not translation or assembly of Solenopsis invicta virus 3 in a baculovirus-driven expression system
Fig. 4. Quantitative PCR (absolute) results evaluating transcript production of SINV-3 by AcSINV-3-infected Sf21 cells. RNA preparations from AcSINV-3 (2 rep- licates, Ci and Di) were treated with DNase I, reverse transcribed, and amplified by qPCR. Results were compared with a series of plasmid constructs containing a portion of the SINV-3 genome (102–109 genome equivalents). The region amplified was at the 3'-most end of ORF2 (see Fig. 2). No amplification was detected in polyhedrin-negative AcRP23.lacZ preparations.
Data from: Baculovirus infection triggers a positive phototactic response in caterpillars (a response to Dobson et al. Biol Letters 2015).
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Data from: Baculovirus-induced tree-top disease: how extended is the role of egt as a gene for the extended phenotype?
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Transcriptome sequencing of and microarray development for a Helicoverpa zea cell line to investigate in vitro insect cell-baculovirus interactions
GEO Series GSE34418. Helicoverpa zea. 4 samples. Type: Expression profiling by array.
MicroRNAome of Spodoptera frugiperda cells (Sf9) and its alteration following baculovirus infection
GEO Series GSE60064. Spodoptera frugiperda. 3 samples. Type: Non-coding RNA profiling by high throughput sequencing.
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Allen Brain Atlas
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DANDI Archive for NWB datasets
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International Brain Laboratory public data
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OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.