About: The Production of Recombinant Infectious DI-Particles of a Murine Coronavirus in the Absence of Helper Virus

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About: The Production of Recombinant Infectious DI-Particles of a Murine Coronavirus in the Absence of Helper Virus Goto Sponge NotDistinct Permalink

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

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	The Production of Recombinant Infectious DI-Particles of a Murine Coronavirus in the Absence of Helper Virus
Creator	Luytjes, Willem Spaan, Willy Bos, Evelyne Koerten, Henk Van Der Meulen, Hans
Source	Elsevier; Medline; PMC
abstract	Abstract We have studied the production and release of infectious DI-particles in vaccinia-T7-polymerase recombinant virus-infected L cells that were transfected with five different plasmids expressing the synthetic DI RNA MIDI-HD and the four structural proteins (M, N, S, and E) of the murine coronavirus MHV-A59. The DI cDNA contains the hepatitis delta ribozyme sequences to generate in the transfected cells a defined 3′ end. In EM studies of transfected cells virus-like particles (VLP) were observed in vesicles. Release of the particles into the medium was studied by immunoprecipitations of proteins released into the culture supernatant. Particle release was independent of S or N, but required M and E. Coexpression of E and M was sufficient for particle release. Coexpression of the structural proteins and the MIDI-HD RNA resulted in the production and release of infectious DI-particles. Infectivity of the DI-particles was determined by adding helper virus MHV-A59 to the medium containing the VLPs and using this mixture to infect new L cells. Intracellular RNA of several subsequent undiluted passages was isolated to detect the MIDI-HD RNA. Passage of the MIDI-HD RNA was dependent on the expression of the structural proteins of MHV-A59 in the transfected cells. In the absence of either E or M, MIDI-HD RNA could not be passaged to fresh L cells. We have thus developed a system in which we can produce coronavirus-like particles and an assay to test their infectivity.
has issue date	1996-04-01(xsd:dateTime)
bibo:doi	10.1006/viro.1996.0165
bibo:pmid	8615041
has license	els-covid
sha1sum (hex)	5b44af2cdde5e0824570443c614a8d52ebeed552
schema:url	https://doi.org/10.1006/viro.1996.0165
resource representing a document's title	The Production of Recombinant Infectious DI-Particles of a Murine Coronavirus in the Absence of Helper Virus
has PubMed Central identifier	PMC7130547
has PubMed identifier	8615041
schema:publication	Virology
resource representing a document's body	covid:5b44af2cdde5e0824570443c614a8d52ebeed552#body_text
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