About: Recombinant Genomic RNA of Coronavirus MHV-A59 after Coreplication with a DI RNA Containing the MHV-RI Spike Gene

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About: Recombinant Genomic RNA of Coronavirus MHV-A59 after Coreplication with a DI RNA Containing the MHV-RI Spike Gene Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	Recombinant Genomic RNA of Coronavirus MHV-A59 after Coreplication with a DI RNA Containing the MHV-RI Spike Gene
Creator	Luytjes, Willem Spaan, Willy Zhang, Linong Homberger, Felix
topic	covid:aa23967b98943923bde1e9a0e2b3470d3966c829#this
source	Elsevier; Medline; PMC
abstract	Abstract A strategy for targeted RNA recombination between the spike gene on the genomic RNA of MHV-A59 and a synthetic DI RNA containing the MHV-RI spike gene is described. The MHV-RI spike gene contains several nucleotide differences from the MHV-A59 spike gene that could be used as genetic markers, including a stretch of 156 additional nucleotides starting at nucleotide 1497. The MHV-RI S gene cDNA (from nucleotide 277-termination codon) was inserted in frame into pMIDI, a full-length cDNA clone of an MHV-A59 DI, yielding pDPRIS. Using the vaccinia vTF7.3 system, RNA was transcribed from pDPRIS upon transfection into MHV-A59-infected L cells. DPRIS RNA was shown to be replicated and passaged efficiently. MHV-A59 and the DPRIS DI particle were copassaged several times. Using a highly specific and sensitive RT-PCR, recombinant genomic RNA was detected in intracellular RNA from total lysates of pDPRIS-transfected and MHV-A59-infected cells and among genomic RNA that was agarose gel-purified from these lysates. More significantly, specific PCR products were found in virion RNA from progeny virus. PCR products were absent in control mixes of intracellular RNA from MHV-A59-infected cells andin vitro-transcribed DPRIS RNA. PCR products from intracellular RNA and virion RNA were cloned and 11 independent clones were sequenced. Crossovers between A59 and RI RNA were found upstream of nucleotide 1497 and had occurred between 106 nucleotides from the 5′-border and 73 nucleotides from the 3′-border of sequence homologous between A59 and RI S genes. We conclude that homologous RNA recombination took place between the genomic RNA template and the synthetic DI RNA template at different locations, generating a series of MHV recombinant genomes with chimeric S genes.
has issue date	1997-03-31(xsd:dateTime)
bibo:doi	10.1006/viro.1997.8460
bibo:pmid	9126265
has license	els-covid
sha1sum (hex)	aa23967b98943923bde1e9a0e2b3470d3966c829
schema:url	https://doi.org/10.1006/viro.1997.8460
resource representing a document's title	Recombinant Genomic RNA of Coronavirus MHV-A59 after Coreplication with a DI RNA Containing the MHV-RI Spike Gene
has PubMed Central identifier	PMC7130785
has PubMed identifier	9126265
schema:publication	Virology
resource representing a document's body	covid:aa23967b98943923bde1e9a0e2b3470d3966c829#body_text
is http://vocab.deri.ie/void#inDataset of	https://covidontheweb.inria.fr:4443/about/id/http/ns.inria.fr/covid19/aa23967b98943923bde1e9a0e2b3470d3966c829
is schema:about of	named entity 'RNA' named entity 'recombinant' named entity 'RNA' named entity 'efficiently' named entity 'gene' named entity 'recombinant' named entity 'products' named entity 'replicated' named entity 'sensitive' named entity 'Spike' named entity 'RNA' named entity 'SPIKE' named entity 'GENE' named entity 'INSERTED' named entity 'TRANSCRIBED' named entity 'HIGHLY' named entity 'MHV' named entity 'RNA' named entity 'CONTAINING' named entity 'COULD BE' named entity 'VIRION' named entity 'ACADEMIC' named entity 'PURIFIED' named entity 'FOUND' named entity 'LENGTH' named entity 'RNA RECOMBINATION' named entity 'VIRUS' named entity 'CDNA CLONE' named entity 'STRETCH' named entity 'INCLUDING' named entity 'CELLS' named entity 'CONTAINS' named entity 'GENE' named entity 'GENES' named entity 'A59' named entity 'A59' named entity 'MHV' named entity 'RT-PCR' named entity 'NUCLEOTIDE' named entity 'USING' named entity 'TIMES' named entity 'SYNTHETIC' named entity 'CONCLUDE' named entity 'BORDER' named entity 'PRODUCTS' named entity 'GENETIC MARKERS' named entity 'TARGETED' named entity 'HOMOLOGOUS' named entity 'PROGENY' named entity 'USED' named entity 'REPLICATED' named entity 'ABSENT' named entity 'SYSTEM' named entity 'AGAROSE GEL' named entity 'CONTROL' named entity 'GENERATING' named entity 'INDEPENDENT' named entity 'RECOMBINANT' named entity 'CORONAVIRUS' named entity 'GENOMIC' named entity 'PCR' named entity 'CLONED' named entity 'FRAME' named entity 'GENOMIC' named entity 'CONTAINING' named entity 'RNA' named entity 'SERIES' named entity 'STARTING' named entity 'STRATEGY' named entity 'SEQUENCE' named entity 'DESCRIBED' named entity '156' named entity 'INFECTED' named entity 'ADDITIONAL' named entity 'RECOMBINANT' named entity 'NUCLEOTIDES'

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