Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.6 (RNA polymerase)
34,946 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The CTN rabies virus was isolated from a human in China in 1953 and subsequently attenuated by multiple passaging to a vaccine strain now approved by the WHO. In this study, we describe the development of a reverse genetics system for the CTN rabies virus strain. The recombinant full-length genomic cDNA was flanked by a hammerhead ribozyme (HamRz) and the hepatitis delta virus ribozyme (HdvRz) while the non-coding G-L region was replaced with a green fluorescent protein (GFP) gene. A set of helper plasmids encoding nucleoprotein (N), phosphoprotein (P), and large protein (L) were constructed and co-transfected with recombinant full-length genome plasmid into BHK-21 cells. Recombinant virus was successfully recovered from cloned cDNA under control of the CMV promoter driven by RNA polymerase II. The recombinant virus, CTN-GFP, stably expressed GFP as detected by fluorescence microscopy. A group of 1-day-old suckling mice was challenged with the CTN-GFP strain by intracerebral inoculation, resulting in 100% morbidity and GFP expression was detected in brain tissues. The recombinant virus CTN-GFP strain recovered from cloned cDNA will be useful as a viral vector to express other foreign genes.
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PMID:Development of a reverse genetics system for a human rabies virus vaccine strain employed in China. 2008 Jan 36

Bats have been identified as a natural reservoir for an increasing number of emerging zoonotic viruses, such as Hendra virus, Nipah virus, Ebola virus, Marburg virus, rabies and other lyssaviruses. Recently, a large number of viruses closely related to members of the genus Coronavirus have been associated with severe acute respiratory syndrome (SARS) and detected in bat species. In this study, samples were collected from 106 live bats of seven different bat species from 27 different locations in Slovenia. Coronaviruses were detected by RT-PCR in 14 out of 36 horseshoe bat (Rhinolophus hipposideros) fecal samples, with 38.8% virus prevalence. Sequence analysis of a 405-nucleotide region of the highly conserved RNA polymerase gene (pol) showed that all coronaviruses detected in this study are genetically closely related, with 99.5-100% nucleotide identity, and belong to group 2 of the coronaviruses. The most closely related virus sequence in GenBank was SARS bat isolate Rp3/2004 (DQ071615) within the SARS-like CoV cluster, sharing 85% nucleotide identity and 95.6% amino acid identity. The potential risk of a new group of bat coronaviruses as a reservoir for human infections is highly suspected, and further molecular epidemiologic studies of these bat coronaviruses are needed.
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PMID:Identification of SARS-like coronaviruses in horseshoe bats (Rhinolophus hipposideros) in Slovenia. 2021 55

An unusual sickness in mules at Batrasi camp, District Mansehra, Pakistan, was reported. Twelve animals died with in 2-3 days after showing the clinical symptoms confusing with colic and nervous disorders. Animals did not respond to any treatment. A team of veterinary doctors/researchers from institute visited the place and collected the samples and information in all aspects related to any disease occurrence on epidemiological basis. Animals were also showing symptoms confusing with rabies. Brain samples were collected for rabies testing. Reverse transcriptase polymerase chain reaction (RT-PCR) by amplifying "N" region gene and mouse inoculation test (MIT) were performed and results showed that disease was nothing except rabies and RT-PCR is the rapid and sensitive method for diagnosis of rabies virus as compared to other conventional methods of diagnosis.
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PMID:Rabies out-break in mules at Mansehra, Pakistan. 2048 43

Rabies, which is an acute, progressive, fatal zoonotic infectious disease, is almost always caused by the bite of rabid animals containing rabies virus in their saliva. Since there is no established specific therapy for rabies, preventive and prophylactic measures are of critical importance. In this report a case of human rabies diagnosed antemortem, was presented. A 29 year old man was admitted to Harran University Hospital (in Sanliurfa province, located at southeastern Anatolia) emergency service with symptoms of high fever, general weakness, paresthesia of the right arm, hypersalivation and dysphagia. The patient with poor socioeconomical status was living in a rural area and his anamnesis revealed a history of dog bite about five months ago. It was learned that he refused vaccination against rabies after the bite event, despite the warnings of his relatives. Shortly after admission, the patient's neurological status severly deteriorated; he became increasingly agitated. Upon the development of progressive respiratory failure, the patient underwent ventilatory support and heavily sedated with presumptive diagnosis of rabies. A nuchal skin biopsy, cerebrospinal fluid, saliva and corneal smear were sent to the Ministry of Agriculture and Rural Affairs Etlik Central Veterinary Control and Research Institute Rabies Diagnosis Laboratory in Ankara. The corneal smear was positive for rabies virus antigen revealed by direct fluorescent antibody test and saliva sample was also positive for rabies virus RNA by reverse-transcriptase polymerase chain reaction assay. Thus, on the third day of the admission the diagnosis was confirmed and on day 11, the patient was deceased due to rabies encephalitis. This case report emphasizes the importance of public education particularly in low socio-economic and socio-cultural areas, about rabies transmission and preventive and prophylactic measures that should be taken after animal bite.
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PMID:[A human rabies case with antemortem diagnosis]. 2054 67

The rescue of recombinant rabies virus (RV) from cloned cDNA is an inefficient process because it relies on the de novo formation within cells of functional ribonucleoprotein (RNP) complexes from plasmid-expressed viral-like antigenome RNAs and three helper proteins. In the standard RV reverse genetics systems, bacteriophage T7 RNA polymerase drives the transcription of virus antigenome-like RNAs containing three nonviral G residues at the 5'-end and a correct 3'-end generated by the autocatalytic activity of an 85 nucleotides long hepatitis delta virus antigenomic "core" ribozyme (HDVagrz). Here, we show that employing optimized ribozyme sequences significantly improves RV rescue. Substitution of the "core" HDVagrz by a ribozyme with an enhanced cleavage activity resulted in an approximately 10-fold higher number of rescue events and faster initiation of an infectious cycle. The alternative use of a hammerhead ribozyme for the generation of an exact 5'-end similarly enhanced rescue efficiency. Notably, RV cDNA clones containing the combination of optimized 3'- and 5'-ribozymes were rescued at an at least 100-fold increase. In addition to virus rescue, reporter gene expression from transfected minigenome cDNAs was significantly enhanced by the novel ribozymes. The improved RV reverse genetics system greatly facilitates recovery of strongly attenuated viruses and vectors for biomedical applications.
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PMID:Significantly improved rescue of rabies virus from cDNA plasmids. 2139 81

The first report of the raccoon variant of rabies virus was in Ontario, Canada in 1999. As part of the control of this outbreak a Point Infection Control (PIC) strategy of trapping and euthanizing vector species was implemented. To evaluate whether this strategy was indeed removing diseased animals, rabies diagnosis was performed on these specimens. During a PIC program conducted in 2003, 721 animals (raccoons, striped skunks and red foxes) were collected and euthanized and brain material from each specimen was divided into two halves; one half was submitted for rabies diagnosis by a direct fluorescent antibody (DFA) test while the other was tested using a sensitive real-time reverse-transcriptase polymerase chain reaction (RT-qPCR), to detect raccoon rabies virus (RRV) RNA. This latter assay can detect less than ten viral copies in 200ng of total cellular RNA. All 721 PIC brain samples were negative by the DFA test but ten of them (5 raccoons, 5 skunks) tested positive for raccoon rabies virus by the RT-qPCR assay albeit at low levels. Three of these samples were confirmed by sequencing of the PCR products. Little correlation was observed between clinical rabies DFA positive scoring categories and viral copy number as determined by RT-qPCR.
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PMID:Re-assessment of direct fluorescent antibody negative brain tissues with a real-time PCR assay to detect the presence of raccoon rabies virus RNA. 2151 25

The transcription mode of rabies virus high egg passage-Flury (HEP) strain was examined and compared with that of Evelyn Rokitniki Abelseth (ERA) strain by northern blot analysis using rabies virus gene-specific probes. The ERA strain was shown to exclusively produce monocistronic mRNAs in transcription. All combinations of multicistronic transcripts, including five monocistronic mRNAs, were detected in the viral RNA transcripts of HEP strain. It was concluded that the unique transcription mode is not due to the nucleotide structure of the genome RNA template, but rather to the viral RNA polymerase of HEP strain. The viral polymerase of HEP strain read through the gene junction at a high frequency. The HEP strain has been passaged many times in chick embryo and cultured cells, and has adapted to propagate well in the baby hamster kidney-21 (BHK-21) cells. Through these passages in various hosts, the HEP strain has acquired a unique transcription mode that might have an advantage in amplification of the virus.
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PMID:A unique transcription mode of rabies virus high egg passage-Flury strain detected in infected baby hamster kidney-21 cells. 2164 51

Arctic foxes, 620 that were trapped and 22 found dead on Svalbard, Norway (1996-2004), as well as 10 foxes trapped in Nenets, North-West Russia (1999), were tested for rabies virus antigen in brain tissue by standard direct fluorescent antibody test. Rabies antigen was found in two foxes from Svalbard and in three from Russia. Blood samples from 515 of the fox carcasses were screened for rabies antibodies with negative result. Our results, together with a previous screening (1980-1989, n=817) indicate that the prevalence of rabies in Svalbard has remained low or that the virus has not been enzootic in the arctic fox population since the first reported outbreak in 1980. Brain tissues from four arctic foxes (one from Svalbard, three from Russia) in which rabies virus antigen was detected were further analyzed by reverse-transcriptase polymerase chain reaction direct amplicon sequencing and phylogenetic analysis. Sequences were compared to corresponding sequences from rabies virus isolates from other arctic regions. The Svalbard isolate and two of the Russian isolates were identical (310 nucleotides), whereas the third Russian isolate differed in six nucleotide positions. However, when translated into amino acid sequences, none of these substitutions produced changes in the amino acid sequence. These findings suggest that the spread of rabies virus to Svalbard was likely due to migration of arctic foxes over sea ice from Russia to Svalbard. Furthermore, when compared to other Arctic rabies virus isolates, a high degree of homology was found, suggesting a high contact rate between arctic fox populations from different arctic regions. The high degree of homology also indicates that other, and more variable, regions of the genome than this part of the nucleoprotein gene should be used to distinguish Arctic rabies virus isolates for epidemiologic purposes.
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PMID:Rabies in the arctic fox population, Svalbard, Norway. 2210 65

This study describes the results of the sequencing and analysis of segments of Blocks II and III of the RNA polymerase L gene of Rabies virus isolates from different reservoir species of Brazil. The phylogenetic relations of the virus were determined and a variety of species-specific nucleotides were found in the analyzed areas, but the majority of these mutations were found to be synonymous. However, an analysis of the putative amino acid sequences were shown to have some characteristic mutations between some reservoir species of Brazil, indicating that there was positive selection in the RNA polymerase L gene of Rabies virus. On comparing the putative viral sequences obtained from the Brazilian isolates and other Lyssavirus, it was determined that amino acid mutations occurred in low-restriction areas. This study of the L gene of Rabies virus is the first to be conducted with samples of virus isolates from Brazil, and the results obtained will help in the determination of the phylogenetic relations of the virus.
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PMID:Phylogenetic analysis of partial RNA-polymerase blocks II and III of Rabies virus isolated from the main rabies reservoirs in Brazil. 2252 40

The aG rabies virus strain has been attenuated through multiple passages in cells and is now used as a vaccine strain in China. We attempted to develop a reverse genetics system using the aG strain. Recombinant full-length genomic cDNA was flanked by a hammerhead ribozyme and the hepatitis delta virus ribozyme. Three helper plasmids encoding the nucleoprotein, the phosphoprotein, and the large protein were produced and introduced together with a plasmid containing the full-length aG viral genome into BHK-21 cells by transfection. Recombinant virus was successfully recovered from the cloned cDNA under the control of a CMV promoter driven by RNA polymerase II. The recombinant virus was confirmed by RT-PCR, and the titer of the recombinant virus was 6.2 log LD50.
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PMID:Development of a reverse genetics system for the aG strain of rabies virus in China. 2427 86


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