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Analysis of secondsite revertants of a murine coronavirus nucleocapsid protein deletion mutant and construction of nucleocapsid protein mutants by targeted RNA recombination (1995)

by D Peng, C A Koetzner, P S Masters, Ding Peng, Cheri A. Koetzner, Paul S. Masters
Venue:Journal of Virology
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Insertion of a New Transcriptional Unit into the Genome of Mouse Hepatitis Virus

by Bilan Hsue, Paul S. Masters, Updated Information, Bilan Hsue, Paul, S. Masters , 1998
"... This article cites 42 articles, 28 of which can be accessed free ..."
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This article cites 42 articles, 28 of which can be accessed free
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...ame gel. Broken lines indicate detectable mobility differences of RNA4 through RNA7 of Al6184 and Al6185. which resulted from separate transfections, were isolated and purified. Direct RNA sequencing =-=(30)-=- of total cytoplasmic RNA derived from Alb169- and Alb170-infected mouse 17 Cl1 cells demonstrated the repair of the Alb4 deletion and the incorporation of IGS7/8 and the adjacent polylinker into each...

A bulged stem-loop structure in the 39 untranslated region of the genome of the coronavirus mouse hepatitis virus is essential for replication

by Bilan Hsue, Paul, S. Masters - J Virol , 1997
"... The 3 * untranslated region (UTR) of the positive-sense RNA genome of the coronavirus mouse hepatitis virus (MHV) contains sequences that are necessary for the synthesis of negative-strand viral RNA as well as sequences that may be crucial for both genomic and subgenomic positive-strand RNA synthesi ..."
Abstract - Cited by 9 (2 self) - Add to MetaCart
The 3 * untranslated region (UTR) of the positive-sense RNA genome of the coronavirus mouse hepatitis virus (MHV) contains sequences that are necessary for the synthesis of negative-strand viral RNA as well as sequences that may be crucial for both genomic and subgenomic positive-strand RNA synthesis. We have found that the entire 3 * UTR of MHV could be replaced by the 3 * UTR of bovine coronavirus (BCV), which diverges overall by 31 % in nucleotide sequence. This exchange between two viruses that are separated by a species barrier was carried out by targeted RNA recombination. Our results define regions of the two 3 * UTRs that are functionally equivalent despite having substantial sequence substitutions, deletions, or insertions with respect to each other. More significantly, our attempts to generate an unallowed substitution of a particular portion of the BCV 3 * UTR for the corresponding region of the MHV 3 * UTR led to the discovery of a bulged stem-loop RNA secondary structure, adjacent to the stop codon of the nucleocapsid gene, that is essential for MHV viral RNA replication. Coronaviruses are a family of single-stranded, positive-po-larity RNA viruses having genomes of 26 to 31 kb that are the largest known replicating RNA molecules (36). Upon infec-tion, coronavirus genomic RNA serves as the message for
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... full genome required for viral replication. Since we have previously shown that it is possible to incorporate mutations into the nucleocapsid (N) gene and the 39 UTR of MHV by targeted recombination =-=(5, 15, 24, 27, 28)-=-, we sought to replace the MHV 39 UTR with a similar sequence by this technique with the aim of revealing critical regions with respect to the MHV replication mechanism. Although the 39 UTRs of differ...

Utilizing fowlpox virus recombinants to generate defective RNAs of the coronavirus infectious bronchitis virus

by Sharon Evans, David Cavanagh, Paul Britton , 2000
"... of the coronavirus infectious bronchitis virus ..."
Abstract - Cited by 6 (5 self) - Add to MetaCart
of the coronavirus infectious bronchitis virus
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...ined for the coronaviruses murine hepatitis virus (MHV) (Fischer et al., 1997, 1998 ; Hsue & Masters, 1997, 1999 ; Koetzner et al., 1992 ; Kuo et al., 2000 ; Liao & Lai, 1992 ; Masters et al., 1994 ; =-=Peng et al., 1995-=-a, b ; Phillips et al., 1999 ; van der Most et al., 1992) and transmissible gastroenteritis virus (TGEV) (Sanchez et al., 1999). We have been developing an IBV-based D-RNA system as an RNA vector for ...

unknown title

by M. H. Verheije, M. V. Kroese, P. J. M. Rottier, J. J. M. Meulenberg
"... Viable porcine arteriviruses with deletions proximal to the 3 � end of the genome ..."
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Viable porcine arteriviruses with deletions proximal to the 3 � end of the genome
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... the best-studied coronavirus, that a 29 amino acid deletion in the putative spacer region preceding the C-terminal domain of the N protein resulted in temperature-sensitive and thermolabile viruses (=-=Peng et al., 1995-=-), we investigated whether our deletion mutants had similar characteristics; they appeared not to have these characteristics. Moreover, infectious virus was still not produced from the deletion mutant...

Characterization of Two Temperature-Sensitive Mutants of Coronavirus Mouse Hepatitis Virus Strain A59 with Maturation

by J. Virol, W Luytjes, H Gerritsma, E Bos, W Spaan, Willem Luytjes, Heleen Gerritsma, Evelyne Bos, Willy Spaan , 1996
"... protein. strain A59 with maturation defects in the spike mutants of coronavirus mouse hepatitis virus Characterization of two temperature-sensitive ..."
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protein. strain A59 with maturation defects in the spike mutants of coronavirus mouse hepatitis virus Characterization of two temperature-sensitive
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...in the context of natural infections, and much remains to be elucidated. Only the nucleocapsid gene is currently subjectable to mutagenesis on the genomic RNA by a targeted RNA recombination protocol =-=(24, 25)-=-. Our current state of knowledge of the synthesis of the MHV structural proteins can be summarized as follows (for recent reviews, see references 4, 15, and 28). Membrane proteins S and M (probably al...

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by Bilan Hsue, Toinette Hartshorne, Paul S. Masters , 2000
"... This article cites 56 articles, 39 of which can be accessed free ..."
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This article cites 56 articles, 39 of which can be accessed free
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...the manufacturer’s instructions. Direct RNA sequencing was performed by a modification of a dideoxy chain termination procedure, using avian myeloblastosis virus reverse transcriptase (Life Sciences) =-=(6, 38)-=-. Radiolabeling of viral RNA and analysis of DI RNA replication. Metabolic labeling of virus-specific RNA was carried out as previously described (12, 32). In brief, L2 cells in spinner culture were i...

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by Philip Hurst, Julie Parsonnet, Sharon Perry, Luz Sanchez, Shufang Yang, Zubin Agarwal
"... This article cites 38 articles, 11 of which can be accessed free ..."
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This article cites 38 articles, 11 of which can be accessed free
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... be reinforced by numerous sequences that later became available. Part of this model, the delineation of spacer B and the acidic, carboxy-terminal domain 3, has been well supported by subsequent work =-=(22, 25, 41, 42)-=-. However, a wealth of recent, detailed structural studies of bacterially expressed domains of the N proteins of the severe acute respiratory syndrome coronavirus (SARS-CoV) and of infectious bronchit...

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by M. H. Verheije, M. V. Kroese, P. J. M. Rottier, J. J. M. Meulenberg , 2016
"... Viable porcine arteriviruses with deletions proximal to the 3 « end of the genome ..."
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Viable porcine arteriviruses with deletions proximal to the 3 « end of the genome
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... the best-studied coronavirus, that a 29 amino acid deletion in the putative spacer region preceding the C-terminal domain of the N protein resulted in temperature-sensitive and thermolabile viruses (=-=Peng et al., 1995-=-), we investigated whether our deletion mutants had similar characteristics ; they appeared not to have these characteristics. Moreover, infectious virus was still not produced from the deletion mutan...

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by Peter J. M. Rottier, Cornelis A. M. De Haan, Lili Kuo, Paul S. Masters, Harry Vennema , 1997
"... This article cites 75 articles, 50 of which can be accessed free at: ..."
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This article cites 75 articles, 50 of which can be accessed free at:
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...IES CORONAVIRUS HOST RANGE MUTANT fMHV 1395so nsFebruary 27, 2014 by PENN STATE UNIV http://jvi.asm.org/ D ow nloaded from Direct RNA sequencing was performed by a modified dideoxy termination method =-=(11, 40)-=-. Intracellular viral protein analysis. LR7 cells and FCWF cells were grown in 35-mm dishes and infected with MHV-A59, fMHV, or FIPV at a multiplicity of 10 PFU per cell. Before being labeled, the cel...

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by J. Virol, Louise Doyon, Catherine Payant, Léa Brakier-gingras , 1997
"... This article cites 21 articles, 11 of which can be accessed free at: ..."
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This article cites 21 articles, 11 of which can be accessed free at:
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...ribed. We have been able to generate site-directed mutations in MHV by targeted recombination between transfected synthetic donor RNA species and recipient mutant viruses that can be selected against =-=(13, 14, 19, 22, 32, 33)-=-. In the present study, this technique was taken a step further in that engineered mutants were identified by screening rather than by * Corresponding author. Mailing address: David Axelrod Institute,...

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