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)

Subgenomic mRNA from a virulent field isolate of porcine transmissible gastroenteritis virus (TGEV), strain FS772/70, was used to produce cDNA. The cDNA from three overlapping clones was sequenced by the chain termination method and two open reading frames (ORFs) were identified. The largest ORF, 4350bp, encoded a polypeptide of 1449 amino acids of relative molecular mass (Mr) 159811, which contained 33 potential N-linked glycosylation sites, a cysteine-rich region, and a potential transmembrane region. The C-terminal half of this ORF showed homology to the S proteins of four other coronaviruses. The other ORF consisted of the 3'-end of a gene with homology to the carboxyl terminus of the F2 subunit of infectious bronchitis virus (IBV) RNA polymerase.
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PMID:Sequence of the S gene from a virulent British field isolate of transmissible gastroenteritis virus. 196 22

The E2-peplomer protein gene of the virulent Miller strain of transmissible gastroenteritis virus (TGEV) was sequenced from cDNA clones and compared to the E2 gene sequence of the avirulent Purdue strain. Sequence comparisons indicate that most amino acid differences occur in the N-terminal half of the E2-peplomer which represents the most exposed region of the protein. In addition, analysis of an incompletely sequenced open reading frame (ORF) to the immediate 5' side of the E2 gene indicates extensive sequence homology with the infectious bronchitis virus (IBV) F2 gene which is thought to encode a RNA polymerase.
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PMID:Nucleotide sequence of the E2-peplomer protein gene and partial nucleotide sequence of the upstream polymerase gene of transmissible gas gastroenteritis virus (Miller strain). 196 16

The nucleotide sequence of the human coronavirus 229E (HCV 229E) RNA polymerase gene and the 5' region of the genome has been determined. The polymerase gene is comprised of two large open reading frames, ORF1a and ORF1b, that contain 4086 and 2687 codons, respectively. ORF1b overlaps ORF1a by 43 bases in the (-1) reading frame. The in vitro translation of SP6 transcripts which include HCV 229E sequences encompassing the ORF1a/ORF1b junction show that expression of ORF1b can be mediated by ribosomal frame-shifting. The predicted translation products of ORF1a (454,200 molecular weight) and ORF1a/1b (754,200 molecular weight) have been compared to the predicted RNA polymerase gene products of infectious bronchitis virus (IBV) and murine hepatitis virus (MHV) and conserved structural features and putative functional domains have been identified. This analysis completes the nucleotide sequence of the HCV 229E genome.
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PMID:Nucleotide sequence of the human coronavirus 229E RNA polymerase locus. 833 38

We have previously reported the nucleotide sequences of gene 2 (spike (S) protein gene), gene 6 (nucleocapsid (N) protein gene), and the 3' end untranslated region of a novel avian infectious bronchitis virus (IBV) strain, CU-T2 [Jia et al. (1995) Arch. Virol. 140, 259 271]. In the present report we describe the sequences of the remaining genes of this strain (gene 3, 4 and 5) with the exception of gene 1 (RNA polymerase gene). Gene 3 contained three open reading frames (ORFs), 3a, 3b and 3c of 174, 195 and 282 nucleotides (nt), respectively. Gene 4 (membrane (M) protein gene) consisted of 749 nt with a single ORF of 687 nt. Gene 5 contained two ORFs, 5a and 5b, with 198 and 249 nt, respectively. Thus, in total, there were 7349 nt from the 5' end of S protein gene to the 3' end of the CU-T2 genome. The overall nt sequence homologies between gene 3, 4, and 5 of CU-T2 and those of other strains were between 84.1-90.8%, 85.8-88.8% and 90.4 96.4%, respectively. The predicted amino acid (aa) sequence homologies revealed that gene 3b and 5b were more conserved than 3a, 3c and 5a. Each individual gene of CU-T2 strain (with the exception of the RNA polymerase gene) had a different level of homology with the homologous gene of other strains, suggesting that the evolution of IBV strains in general has been a complex, and as yet, poorly understood process.
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PMID:Sequence analysis of gene 3, gene 4 and gene 5 of avian infectious bronchitis virus strain CU-T2. 916 26

Coronavirus defective RNAs (D-RNAs) have been used as RNA vectors for the expression of heterologous genes and as vehicles for reverse genetics by modifying coronavirus genomes by targetted recombination. D-RNAs based on the avian coronavirus infectious bronchitis virus (IBV) D-RNA CD-61 have been rescued (replicated and packaged into virions) in a helper virus-dependent manner following electroporation of in vitro-generated T7 transcripts into IBV-infected cells. In order to increase the efficiency of rescue of IBV D-RNAs, cDNAs based on CD-61, under the control of a T7 promoter, were integrated into the fowlpox virus (FPV) genome. The 3'-UTR of the D-RNAs was flanked by a hepatitis delta antigenomic ribozyme and T7 terminator sequence to generate suitable 3' ends for rescue by helper IBV. Cells were co-infected simultaneously with IBV, the recombinant FPV (rFPV) containing the D-RNA sequence and a second rFPV expressing T7 RNA polymerase for the initial expression of the D-RNA transcript, subsequently rescued by helper IBV. Rescue of rFPV-derived CD-61 occurred earlier and with higher efficiency than demonstrated previously for electroporation of in vitro T7-generated RNA transcripts in avian cells. Rescue of CD-61 was also demonstrated for the first time in mammalian cells. The rescue of rFPV-derived CD-61 by M41 helper IBV resulted in leader switching, in which the Beaudette-type leader sequence on CD-61 was replaced with the M41 leader sequence, confirming that helper IBV virus replicated the rFPV-derived D-RNA. An rFPV-derived D-RNA containing the luciferase gene under the control of an IBV transcription-associated sequence was also rescued and expressed luciferase on serial passage.
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PMID:Utilizing fowlpox virus recombinants to generate defective RNAs of the coronavirus infectious bronchitis virus. 1108 16

Major advances in the study of the molecular biology of RNA viruses have resulted from the ability to generate and manipulate full-length genomic cDNAs of the viral genomes with the subsequent synthesis of infectious RNA for the generation of recombinant viruses. Coronaviruses have the largest RNA virus genomes and, together with genetic instability of some cDNA sequences in Escherichia coli, this has hampered the generation of a reverse-genetics system for this group of viruses. In this report, we describe the assembly of a full-length cDNA from the positive-sense genomic RNA of the avian coronavirus, infectious bronchitis virus (IBV), an important poultry pathogen. The IBV genomic cDNA was assembled immediately downstream of a T7 RNA polymerase promoter by in vitro ligation and cloned directly into the vaccinia virus genome. Infectious IBV RNA was generated in situ after the transfection of restricted recombinant vaccinia virus DNA into primary chick kidney cells previously infected with a recombinant fowlpox virus expressing T7 RNA polymerase. Recombinant IBV, containing two marker mutations, was recovered from the transfected cells. These results describe a reverse-genetics system for studying the molecular biology of IBV and establish a paradigm for generating genetically defined vaccines for IBV.
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PMID:Reverse genetics system for the avian coronavirus infectious bronchitis virus. 1171 26

In a case-control study of the infectious agents associated with natural outbreaks of respiratory disease in pheasants, 28 batches of birds from sites affected by disease and eight batches of birds from unaffected sites were examined by six veterinary laboratories in England, Wales and Scotland, and tested for mycoplasmas, other bacteria and viruses. Sinusitis was the commonest sign of disease and was associated with Mycoplasma gallisepticum as detected by PCR in the trachea (P < 0.05) and conjunctiva (P < 0.01). Sinusitis was also associated with pasteurella cultured from the sinus (P < 0.05), antibody to avian pneumovirus (APV) (P < 0.01) and avian coronaviruses as detected by reverse-transcriptase PCR (P < 0.05); there was no association between disease and APV as detected by PCR. Avian coronaviruses were the most common infectious agents detected. They were genetically close to infectious bronchitis virus (IBV) but differed in their gene sequence from all the serotypes of IBV previously identified in domestic fowl, and serological tests with six known IBV types showed little cross reactivity. Mycoplasma species other than M gallisepticum were cultured in 18 batches of pheasants but, with the exception of Mycoplasma gallinaceum, were not associated with disease.
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PMID:Infectious agents associated with respiratory disease in pheasants. 1205 35

The entire crop of 18,120 pheasants for the 2000 rearing season (May 8 to August 7) of one estate in the south of England was vaccinated at one day and five weeks of age with a turkey rhinotracheitis (TRT) vaccine. Blood samples and oropharyngeal swabs were taken from the second week's hatching every three weeks throughout the growing season to assess the response of the birds. There was evidence of seroconversion in samples collected three weeks after vaccination, with positive titres being maintained in 33 per cent or more of the population up to at least 22 weeks of age. Positive titres were also recorded in samples taken on December 6 from shot birds between 22 and 30 weeks of age. Positive titres to infectious bronchitis virus (IBV) were identified in a high proportion of the poults as early as one day of age. Reverse-transcriptase PCR detected IBV-like virus and TRT of the same subtype as the TRT vaccine administered three weeks previously.
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PMID:Response of pheasants to live attenuated turkey rhinotracheitis vaccine. 1237 89

Reverse-transcriptase polymerase chain reactions (RT-PCRs) were used to examine RNA extracted from mouth/nasal swabs from pheasants exhibiting signs of respiratory disease. The oligonucleotides used were based on sequences of infectious bronchitis virus (IBV), the coronavirus of domestic fowl. A RT-PCR for the highly conserved region II of the 3' untranslated region of the IBV genome detected a coronavirus in swabs from 18/21 estates. Sequence identity with the corresponding region of IBVs and coronaviruses from turkeys was > 95%. A RT-PCR for part of the S1 region of the spike protein gene was positive with 13/21 of the samples. Sequence analysis of the RT-PCR products derived from nine of the pheasant viruses revealed that some of the viruses differed from each other by approximately 24%, similar to the degree of difference exhibited by different serotypes of IBV. Further analysis of the genome of one of the viruses revealed that it contained genes 3 and 5 that are typical of IBV but absent in both the transmissible gastroenteritis virus and murine hepatitis virus groups of mammalian coronaviruses. The nucleotide sequences of genes 3 and 5 of the pheasant virus had a similar degree of identity (approximately 90%) with those of coronaviruses from turkeys and chickens, as is observed when different serotypes of IBV are compared. This work: (a) confirms that coronaviruses are present in pheasants (indeed, commonly present in pheasants with respiratory disease); (b) demonstrates that their genomes are IBV-like in their organization; and (c) shows that there is sequence heterogeneity within the group of pheasant coronaviruses, especially within the spike protein gene. Furthermore, the gene sequences of the pheasant viruses differed from those of IBV to similar extents as the sequence of one serotype of IBV differs from another. On the genetic evidence to date, there is a remarkably high degree of genetic similarity between the coronaviruses of chickens, turkeys and pheasants.
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PMID:Coronaviruses from pheasants (Phasianus colchicus) are genetically closely related to coronaviruses of domestic fowl (infectious bronchitis virus) and turkeys. 1242 95

The 2 groups of human coronaviruses (HCoVs) represented by the prototype strains HCoV 229E and HCoV OC43 are mostly known as viruses responsible for common cold syndrome. HCoVs are difficult to detect, and epidemiological data are rare. From October 2000 through April 2001, we tested 1803 respiratory samples for HCoV by reverse-transcriptase polymerase chain reaction. From 8 February through 27 March 2001, HCoV OC43 was detected in samples obtained from 30 (6%) of 501 patients. The other viruses detected were respiratory syncytial virus (6.1%), parainfluenza virus 3 (1%), influenza virus A (7.8%), influenza virus B (7.2%), rhinovirus (6.4%), enterovirus (1%), and adenovirus (2%). Infection with HCoV OC43 was detected in patients of all age groups. The following clinical symptoms were noted: fever (in 59.8% of patients), general symptoms (in 30%), digestive problems (in 56.8%), rhinitis (in 36.6%), pharyngitis (in 30%), laryngitis (in 3.3%), otitis (in 13.3%), bronchitis (in 16.6%), bronchiolitis (in 10%), and pneumonia (in 6.6%). This study shows that an outbreak of HCoV OC43 respiratory infection was responsible for the lower respiratory tract symptoms observed in nearly one-third of patients identified by active surveillance for coronavirus infection.
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PMID:An outbreak of coronavirus OC43 respiratory infection in Normandy, France. 1268 10


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