NOVEL METHODS FOR RESCUE OF RNA VIRUSES

摘要

This invention relates to improved methods for producing nonsegmented, negative-sense, single-stranded RNA viruses of the Order designated Mononegavirales virus, including embodiments relating to methods of producin g such viruses as attenuated and/or infectious viruses, such as Measles virus (MV) and respiratory syncytial virus (RSV). One method for producing a recombinant virus from the Order Mononegavirales comprises (a) in at least o ne host cell, conducting transfection of a rescue composition which comprises ( i) a transcription vector comprising an isolated nucleic acid molecule which comprises polynucleotide sequence encoding a genome or antigenome of a nonsegmented, negative-sense, single stranded RNA virus of the Order Mononegavirales and (ii) at least one expression vector which comprises at least one isolated nucleic acid molecule encoding the trans-acting proteins necessary for encapsidation, transcription and replication; in a host cell under conditions sufficient to permit the co-expression of these vectors and the production of the recombinant virus; (b) heating the transfected rescue composition to an effective heat shock temperature under conditions sufficie nt to increase the recovery of the recombinant virus; and optionally, (c) harvesting the resulting recombinant virus.