Infectious clone for human parainfluenza virus type 3

A method of introducing a site-specific mutation into the genome of a recombinant human parainfluenza virus, comprising the following steps:

• a. preparing a clone comprising a nucleotide sequence encoding a human parainfluenza viral antigenome having a mutation at a specifc site;
• b. co-transfecting host cells with the clone of step (a), a support clone comprising a nucleotide sequence encoding an HPIV L protein, and a support clone comprising a nucleotide sequence encoding an HPIV P protein;
• wherein the nucleotide sequence encoding the P protein and L protein may be on the same support clone or on separate support clones;
• and
• c. culturing the transfected host cells for a time sufficient to allow formation of a recombinant human parainfluenza virus.

A host cell for producing a recombinant human parainfluenza type 3 virus, said host cell comprising:

• a. an HPIV-3 clone comprising a nucleotide sequence encoding a positive sense, antigenome of human parainfluenza type 3 virus;
• b. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus type 3 L protein;
• and
• c. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus type 3 P protein;
• wherein the nucleotide sequence encoding the P protein and L protein may be on the same support clone or on separate support clones.

A host cell for producing a recombinant human parainfluenza virus type 3, said host cell comprising:

• a. an HPIV clone comprising a nucleotide sequence encoding a positive sense, antigenome of human parainfluenza virus type 3, wherein said antigenome sequence comprises a site-specific mutation;
• b. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus type 3 L protein;
• and
• c. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus type 3 P protein;
• wherein the nucleotide sequence encoding the P protein and the nucleotide sequence encoding the L protein are on the same support clone or on separate support clones.

A host cell for producing a recombinant human parainfluenza virus, said host cell comprising:

• a. an HPIV clone comprising a nucleotide sequence encoding a positive sense, antigenome of human parainfluenza virus, wherein said antigenome sequence comprises a site specific mutation;
• b. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus L protein;
• and
• c. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus P protein;
• wherein the nucleotide sequences encoding the P protein and the L protein are on the same support clone or on separate support clones;
• and
• wherein the mutation is in the L protein coding region of the antigenome encoding sequence of the HPIV clone, and wherein the support clone which comprises the L protein encoding sequence has a corresponding mutation therein.

A host cell for producing a recombinant human parainfluenza virus, said host cell comprising:

• a. an HPIV clone comprising a nucleotide sequence encoding a positive sense, antigenome of human parainfluenza virus, wherein said antigenome sequence comprises a site specific mutation;
• b. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus L protein;
• and
• c. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus P protein;
• wherein the nucleotide sequences encoding the P protein and the L protein are on the same support clone or on separate support clones;
• and
• wherein the mutation is in the P protein coding region of the antigenome encoding sequence of the HPIV clone, and wherein the support clone which comprises the P protein encoding sequence has a corresponding mutation therein.

A host cell for producing a recombinant human parainfluenza virus, said host cell comprising:

• a. an HPIV clone comprising a nucleotide sequence encoding a positive sense, antigenome of human parainfluenza virus, wherein said antigenome sequence comprises a site specific mutation;
• b. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus L protein;
• c. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus P protein;
• and
• d. a support clone comprising a nucleotide sequence encoding a human parainfluenza virus NP protein;
• wherein the nucleotide sequences encoding the P protein, L protein, and NP protein are on the same support clone or on separate support clones and
• wherein the mutation is in the NP protein coding region of the antigenome encoding sequence of the HPIV clone, and wherein the support clone which comprises the NP protein encoding sequence has a corresponding mutation therein.