PMID: 8615041Apr 1, 1996Paper

The production of recombinant infectious DI-particles of a murine coronavirus in the absence of helper virus

Virology
E C BosW J Spaan

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 ...Continue Reading

Citations

Feb 16, 2010·The Journal of Biological Chemistry·Ying-Tzu TsengChin-Tien Wang
Aug 20, 2010·Journal of Virology·Ariel L ArndtBrenda G Hogue
Jan 19, 2007·Journal of Virology·Ye Ye, Brenda G Hogue
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