Product of infectious respiratory syncytial virus from cloned nucleotide sequences

DETAILED DESCRIPTION The present invention provides an isolated infectious RSV particle which comprises a recombinant RSV genome or antigenome, a nucleocapsid (N) protein, a nucleocapsid phosphoprotein (P), a large (L) polymerase protein, and an RNA polymerase elongation factor.
The RNA polymerase elongation factor can be M2(ORF1) of RSV.
The isolated infectious RSV particle can be a viral or subviral particle.
The isolated infectious RSV virus may be a human RSV, a bovine or murine RSV, or the genome or antigenome can be a chimera of two or more different RSV genomes, such as having nucleotide segments from human and bovine RSV.
In other embodiments the invention provides a method for producing an infectious RSV particle from one or more isolated polynucleotide molecules encoding an RSV.
An expression vector which comprises an isolated polynucleotide molecule encoding a RSV genome or antigenome and an expression vector which comprises one or more isolated polynucleotide molecules that encodes N, P, L and RNA polymerase elongation factor proteins are coexpressed in a cell or cell-free lysate, thereby producing an infectious RSV particle.
The RSV genome or antigenome and the N, P, L and RNA polymerase elongation factor proteins can be coexpressed by the same or different expression vectors.
In some instances the N, P, L and RNA polymerase elongation factor proteins are each encoded on different expression vectors.
The polynucleotide molecule encoding the RSV genome or antigenome is from a human, bovine or murine RSV sequence, and can be a chimera of a human RSV strain sequence and at least one non-human RSV sequence, or can encodes the genome or antigenome of a wild-type RSV strain.
The RSV genome or antigenome can be modified from a wild-type RSV strain by a nucleotide insertion, rearrangement, deletion or substitution, so as to encode a phenotypic alteration such as one that results in attenuation, temperature-sensitivity, cold-adaptation, small plaque size, host range restriction, or a change in an immunogenic epitope of RSV