Production of Red-spotted Grouper Nervous Necrosis Virus (RGNNV) Capsid Protein Using Saccharomyces cerevisiae Surface Display ;kpubs;kpubs.org

ABSTRACT: Betanodaviruses, the causative agents of viral nervous necrosis in marine fish, have bipartite positive-sense RNA genomes. The viruses have been classified into 4 distinct types based on nucleotide sequence similarities in the variable region (the so-called T4 region) of the smaller genomic segment RNA2 (1.4 kb). Betanodaviruses have marked host specificity, although the primary structures of the viral RNAs and encoded proteins are similar among the viruses. We have previously demonstrated, using reassortants between striped jack nervous necrosis virus (SJNNV) and redspotted grouper nervous necrosis virus (RGNNV), that RNA2, which encodes the coat protein, strictly controls host specificity. However, because RNA2 is large, we were unable to propose a mechanism underlying this RNA2-based host specificity. To identify the RNA2 region that controls host specificity, we constructed RNA2 chimeric viruses from SJNNV and RGNNV and tested their infectivity in the original host fish, striped ...

Localization of RNA replication to intracellular membranes is a universal feature of positive-strand RNA viruses. The betanodavirus greasy grouper (Epinephelus tauvina) nervous necrosis virus (GGNNV) is a positive-RNA virus with one of the smallest genomes among RNA viruses replicating in fish cells. To understand the localization of GGNNV replication complexes, we generated polyclonal antisera against protein A, the GGNNV RNA-dependent RNA polymerase. Protein A was detected at 5 h postinfection in infected sea bass cells. Biochemical fractionation experiments revealed that GGNNV protein A sedimented with intracellular membranes upon treatment with an alkaline pH and a high salt concentration, indicating that GGNNV protein A is tightly associated with intracellular membranes in infected cells. Confocal immunofluorescence microscopy and bromo-UTP incorporation studies identified mitochondria as the intracellular site of protein A localization and viral RNA synthesis. In addition, protein A fused ...

Nodaviridae is a family of non-enveloped isometric viruses with bipartite positive-sense RNA genomes. The Nodaviridae family consists of two genera: alpha- and beta-nodavirus. Alphanodaviruses usually infect insect cells. Some commercially available insect cell lines have been latently infected by Alphanodaviruses. A non-enveloped small virus of approximately 30 nm in diameter was discovered co-existing with a recombinant Helicoverpa armigera single nucleopolyhedrovirus (Hear NPV) in Hz-AM1 cells. Genome sequencing and phylogenetic assays indicate that this novel virus belongs to the genus of alphanodavirus in the family Nodaviridae and was designated HzNV. HzNV possesses a RNA genome that contains two segments. RNA1 is 3038 nt long and encodes a 110 kDa viral protein termed protein A. The 1404 nt long RNA2 encodes a 44 kDa protein, which exhibits a high homology with coat protein precursors of other alphanodaviruses. HzNV virions were located in the cytoplasm, in association with cytoplasmic membrane

Viruses in Betanodavirus are non-enveloped, with icosahedral geometries, and T=3 symmetry. The diameter is around 30 nm. Genomes are linear and segmented, bipartite, around 21.4kb in length.[8]. The crystal structure of a betanodavirus- T=3 Grouper nervous necrosis virus (GNNV)-like particle has been determined by X-ray crystallography. The virus-like particle contains 180 subunits of the capsid protein, and each capsid protein (CP) shows three major domains: (i) the N-terminal arm, an inter-subunit extension at the inner surface; (ii) the shell domain (S-domain), a jelly-roll structure; and (iii) the protrusion domain (P-domain) formed by three-fold trimeric protrusions. [10]. ...

SWISS-MODEL Template Library (SMTL) entry for 4rft.1. T=1 subviral particle of Grouper nervous necrosis virus capsid protein deletion mutant (delta 1-34 & 218-338)

The flockhouse virus of the Nodaviridae (bipartitite single stranded RNA genome of 3.1 and 1.4 kb) is pathogenic to beetles in New Zealand, can grow vigorously in mammalian as well as drosophila cells, and can infect plants. See Selling, Allison and Kaesberg in PNAS (1990) Genomic RNA of an insect virus directs synthesis of infectious virions in plants ...

1NOV: Resolution of space-group ambiguity and structure determination of nodamura virus to 3.3 A resolution from pseudo-R32 (monoclinic) crystals.

A protein unlike any other appears to be partially responsible for upsetting the stomachs of the most common animal on the planet.. Results of a Rice University study of the Orsay virus unique to nematodes, the worms that make up 80 percent of living creatures on Earth, have revealed the molecular structure of a fusion protein that forms unusual fibers and attaches the virus to cells and infects them.. The researchers led by structural biologist Yizhi Jane Tao and geneticist Weiwei Zhong said the protein is the first pentameric fiber with potential antiviral applications. They hope it will guide bioengineers as they develop synthetic variations of the virus that can infect new worm hosts, including the parasitic ones that infect humans and animals.. Their research is detailed in the open-access journal PLOS Pathogens.. The Orsay virus, the first known to infect Caenorhabditis elegans, was discovered in France in 2011. It doesnt kill the worm, but infects its digestive tract. Three years ago, ...

The ab initio method with NCSA and phase extension has been successfully implemented to determine the structure of the T = 3 GNNV icosahedral VLP. The key to success was the similarity of the mask (or the envelope) to the target structure. Despite significant variations in the main chains of the CP between FHV and GNNV, the envelopes of these T = 3 capsids are somewhat similar to each other, especially in the spatial position of the P-domain at the quasi-threefold axis. Considering that the P-domain of FHV is much smaller than that of GNNV, the radius of the mask was typically increased to 13 Å to cover the entire structure of GNNV. Other trials using the large spherical-shell mask, which was cut by the bisector planes between neighbouring virus particles, were not successful in phasing. On inspection of this spherical-shell mask, we found that it was so small, covering only half of the P-domain, as to result in a negative effect on the phasing. Tests investigating the simulation trials using ...

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The monoclonal antibody (Mab) against Nodavirus (also know as VNN) is specific for this virus. The MAb has been tested by ELISA with Betanodavirus isolated from four different fish species, including European Sea Bass, Atlantic cod, halibut and Striped Jack.The Mab reacted with Nodavirus isolated from European Sea Bass that has been shown to belong to the RGNNV (red spotted groupe rnervous necrosis virus) cluster. It also reacted against Nodavirus isolated from Atlantic cod and halibut that have been shown to belong to the BFNNV (Barfin flounder nervous necrosis virus) cluster. The Mab did not react with SJNNV (striped jack nervous necrosis virus). SJNNV has been shown to be phylogenetically quite different to the other Nodavirus clusters.. ...

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Molecular Immunology ; Volume 54. p. 443-452. 2013. Krasnov, Aleksei; Kileng, Øyvind; Skugor, Stanko; Jørgensen, Sven Martin; Afanasyev, Sergey; Timmerhaus, Gerrit; Sommer, Ann-Inger; Jensen, Ingvill. ...

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In the present study, we evaluated four commonly used housekeeping genes, viz., actin-β, elongation factor-1α (EF1α), acidic ribosomal protein (ARP), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as internal references for quantitative analysis of immune genes in nervous necrosis virus (NNV)-infected seven-band grouper, Hyporthodus septemfasciatus. Expression profiles of the four genes were estimated in 12 tissues of healthy and infected seven-band grouper. Expression stability of the genes was calculated using the delta Ct method, BestKeeper, NormFinder, and geNorm algorithms. Consensus ranking was performed using RefFinder, and statistical analysis was done using GraphpadPrism 5.0. Tissue-specific variations were observed in the four tested housekeeping genes of healthy and NNV-infected seven-band grouper. Fold change calculation for interferon-1 and Mx expression using the four housekeeping genes as internal references presented varied profiles for each tissue. EF1α and actin-β was the

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POSTDOCTORAL POSITIONS IN VIRAL RNA REPLICATION, GENE EXPRESSION AND VIRUS-HOST INTERACTIONS Creative, highly motivated researchers are sought for postdoctoral positions studying the functions and interactions of viral and host factors in RNA replication, gene expression, and related events in positive-strand RNA viruses. Unique opportunities to identify and study relevant host factors are available through our demonstration that bromovirus and nodavirus RNA replicons can replicate and express foreign genes in yeast, allowing the use of powerful yeast genetics to identify pertinent host genes and to greatly facilitate other experiments (Janda and Ahlquist [1993] Cell 72:961-970; Quadt et al. [1995] PNAS 92:4892-4896; Price et al. (1996) PNAS 93:9465-9470). Bromoviruses and nodaviruses are well-studied viruses with excellent genetic and biochemical resources for in vivo and in vitro studies of fundamental replication mechanisms. Significant opportunities are available in several areas of ...

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The spread of COVID-19 in Wuhan was impeded by harsh government-imposed lockdown measures, noted Dr. Amesh Adalja, a senior scholar with the Johns Hopkins Center for Health Security in Baltimore.. Because of that, its hard to compare Wuhans experience to that of U.S. cities with less restrictive lockdown measures and higher infection rates, he said.. In cities that had aggressive measures in place akin to Wuhan, vaccination would remain the solution to protecting the population from a resurgence of cases, Adalja said. The low prevalence in Wuhan is not applicable to other cities which have reported much higher seroprevalence.. The survey of more than 9,000 Wuhan residents revealed that just under 7% of the citys population carried COVID-19 antibodies by April 2020, the researchers reported.. About 40% of those had neutralizing antibodies to COVID, and follow-up blood sampling found that these antibodies remained at stable levels for at least nine months, the study found.. People with ...

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With the flock house virus (FHV) capsid protein as a vector (FHV RNA 2),the epitopes corresponding to the cleavage regions of rotavirus SA11 Vp4 primed broadly cross reactive neutralizing antibodies in guinea pigs. The epitopes of SA11 Vp4:REA(223～242aa),REB(243～262aa),REC(234～251aa),and their combination REA+REB+REC were expressed on the chimeric protein surface in the recombinant pET FHV RE system,and used to immunize guinea pigs to prepare antisera. The antisera were proven to have strong cross recognizing and cross neutralizing activities to homologous SA11 virus and heterologous Wa,S2,DS 1,Ito Sty 3, St. Thomase, Gottfried, UK, and B223 virus strains. The results suggested that the amino acid sequences of rotavirus Vp4 cleavage region have important immunological functions,and when displayed in the proper positions on the FHV RNA 2 protein vector, these epitopes could be prospective as a new type of epitope based subunit vaccines.

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Total number of confirmed cases now 900+, 26 dead.China restricts travel for 40+ million people as the death toll surges.Two deaths have been reported outside Wuhan.Some residents displaying symptoms are being turned away from hospitals.Hospitals in Wuhan make urgent pleas for help and supplies.Beijing orders PLA medics to assist in Wuhan treating patientsUK and US…

Getting drugs into the brain has proved to be a nanoscale puzzle: Anything bigger than 64 nanometers - about the size of a small virus - gets stuck in the space between brain cells once it gets through the blood-brain barrier. Justin Hanes of Johns Hopkins University School of Medicine and colleagues got around this rule by coating particles destined for brain cells in a dense layer of a polymer called polyethylene glycol.

A genomic analysis of Klosneuviruses suggests that they evolved from small viruses that accumulated genetic material over time, but not all virologists are convinced. 0 Comments. ...

A genomic analysis of Klosneuviruses suggests that they evolved from small viruses that accumulated genetic material over time, but not all virologists are convinced. 0 Comments. ...

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Gain-of-function research created novel SARS-CoV-2 virus In April of this year, the Daily Mail of London broke an incredible story. The Daily Mail reported that the U.S. National Institute of Health issued a $3.7 million grant to research coronaviruses at the Wuhan Institute of Virology. This was shortly after the first cases of covid-19 were being reported in the U.S. It was later reported in Newsweek that the U.S. Agency for International Development was funding the Wuhan Institute of Virology

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In Fall 2016, Loki seemed to gain weight sufficiently. His dietary food was, one at a time, taken off the schedule. His doctor decided to stop it altogether and see what happened. And Loki did and does well. He still needs a few extra small meals to ensure plenty calorie intake, but he does it all with regular food! In addition, increasingly we look at him and wonder how he finished his plate without us asking to take a bite, and another and another. And then yesterday (April 7th, 2017) something huge happened: Loki took his last 1 millilitre of motilium. Completely against all odds and expectations, after trying many times before, we were able to phase out that medicine. Somehow, at the age of 8,5, his stomach decided to start doing what it never was able to do: empty without the support of medicine. This moment is the inspiration to write a post. We want to share how in awe we are with this amazing boy ...

In Fall 2016, Loki seemed to gain weight sufficiently. His dietary food was, one at a time, taken off the schedule. His doctor decided to stop it altogether and see what happened. And Loki did and does well. He still needs a few extra small meals to ensure plenty calorie intake, but he does it all with regular food! In addition, increasingly we look at him and wonder how he finished his plate without us asking to take a bite, and another and another. And then yesterday (April 7th, 2017) something huge happened: Loki took his last 1 millilitre of motilium. Completely against all odds and expectations, after trying many times before, we were able to phase out that medicine. Somehow, at the age of 8,5, his stomach decided to start doing what it never was able to do: empty without the support of medicine. This moment is the inspiration to write a post. We want to share how in awe we are with this amazing boy ...

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This Is The Corona Virus Novel Form Or Corona Virus Causes Wuhan PneumoniaThe Corona virus first infected humans around the middle of the 1960s. This virus is called the Corona virus because the top surface resembles a crown, has the zoonotic natu...

The former director of the Centers for Disease Control and Prevention believes the virus that causes COVID-19 escaped from a lab in Wuhan, China, according to a new interview.

On January 9, the World Health Organization notified the public of a flu-like outbreak in China: a cluster of pneumonia cases had been reported in Wuhan, possibly from vendors exposure to live animals at the Huanan Seafood Market. The US Centers for Disease Control and Prevention had gotten the ...

At a large food market in the Chinese city of Wuhan, signs forbid the selling of wild animals and live fowl, while announcements calling for victory over COVID-19 play on a loop from speakers.

SARS-CoV-2 virus was introduced into the greater Wuhan population by an infected lab worker or animal. These claims from this point on will be referred to as

Chaolong Wang & Xihong Lin: Our COVID-19 transmission dynamics modelling paper in Nature reveals that multifaceted interventions effectively controlled the outbreak in Wuhan, and highlights that substantial covert infections may pose risks of resurgence if reopening without control measures.

Whether you are looking for something to do with the family or with kids, or just for a good day trip, Wuhan has a lot of furry (or scaly) attractions. These are three of the best places to if you are an animal lover, or if you know someone who is.

A newly-unearthed video shows live bats being held in cages at the Wuhan Institute of Virology, a fact omitted by the World Health Organization (WHO) report and apparently denied in

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Kevin McGoff gave a talk at Orsay on his work on the theory of entropy srtuctures and symbolic extensions. This theory was founded by Mike Boyle and Tomasz Downarowicz, among others. This theory relates the continuity properties of the measure-theoretic entropy function with the existence of symbolic topological extensions with measure-theoretic entropies as close as…