Rescue of mumps virus from cDNA
View Patent ↗This invention relates to a method for recombinantly producing, via rescue of mumps virus, a nonsegmented, negative-sense, single-stranded RNA virus, and immunogenic compositions formed therefrom. Additional embodiments relate to methods of producing the mumps virus as an attenuated and/or infectious virus. The recombinant viruses are prepared from cDNA clones, and, accordingly, viruses having defined changes, including nucleotide/polynucleotide deletions, insertions, substitutions and re-arrangements, in the place of the genome are obtained.
1. A method for producing a recombinant mumps virus comprising;
in at least one host cell, conducting transfection or transformation, in media, of a rescue composition which comprises (i) a transcription vector comprising an isolated nucleic acid molecule which comprises a polynucleotide sequence encoding an antigenome of mumps virus, or variant polynucleotide sequence thereof, and (ii) at least one expression vector which comprises one more isolated nucleic acid molecule(s) comprising a polynucleotide sequence encoding the trans-acting proteins (NP, P and L) necessary for encapsidation, transcription and replication; under conditions sufficient to permit the co-expression of said vectors and the production of the recombinant virus;
wherein the isolated nucleic acid molecule encoding an antigenome of mumps virus comprises the polynucleotide sequence selected from the group consisting of SEQ. ID NOS. 1, 11 and 12;
and wherein the recombinant virus is harvested.