Newcastle disease virus infectious clones, vaccines and diagnostic assays
AssigneeID-Lelystad, Instituut voor Dierhouderij en Diergezondheid B.V.
InventorsBernardus Petrus Hubertus Peeters, Olav Sven de Leeuw, Arnoud Leonard Josef Gielkens
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Abstract
The invention relates to the process for generating infectious Newcastle disease virus (NDV) entirely from cloned full-length cDNA and to the use of vaccines and diagnostic assays generated with and derived from the process. The process offers the possibility to modify the NDV genome by means of genetic modification and allows the introduction of mutations, deletions and/or insertions. The process can be used to modify the antigenic makeup of NDV, thus allowing the generation of live NDV marker vaccines which can be serologically distinguished from NDV field strains.
Claims42
An avian-paramyxovirus eDNA at least comprising a nucleic acid sequence corresponding to the 5′-terminal end, as set forth in SEQ ID NO:82, and, corresponding to the 3′-terminal end, as set forth in SEQ ID NO:80 or SEQ ID NO:81, of the genome of avian-paramyxovirus, said nucleic acid sequence allowing generation of an infectious copy of avian-paramyxovirus, wherein said avian-paramyxovirus is selected from the group consisting of avian paramyxovirus type-1, avian paramyxovirus type-2, avian paramyxovirus type-3, avian paramyxovirus type-4, avian paramyxovirus type-5, avian paramyxovirus type-6, avian paramyxovirus type-7, avian paramyxovirus type-8 and avian paramyxovirus type-9.
A cDNA at least comprising a nucleic acid sequence corresponding to the 5′-terminal end, as set forth in SEQ ID NO:82, and, corresponding to the 3′-terminal end, as set forth in SEQ ID NO:80 or SEQ ID NO:81, of the genome of avian-paramyxovirus, said nucleic acid sequence allowing generation of a replicating avian-paramyxovirus minigenome, wherein said avian-paramyxovirus is selected from the group consisting of avian paramyxovirus type-1, avian paramyxovirus type-2, avian paramyxovirus type-3, avian paramyxovirus type-4, avian paramyxovirus type-5, avian paramyxovirus type-6, avian paramyxovirus type-7, avian paramyxovirus type-8 and avian paramyxovirus type-9.
An infectious copy avian-paramyxovirus obtained by a method, said method comprising:
- transfecting at least one cell with cDNA, said cDNA comprising an avian-paramyxovirus cDNA at least comprising a nucleic acid sequence corresponding to the 5′-terminal end, as set forth in SEQ ID NO:82, and, corresponding to the 3′-terminal end, as set forth in SEQ ID NO:80 or SEQ ID NO:81, of the genome of avian-paramyxovirus;
- and
- expressing in said cell an infectious copy of avian-paramyxovirus.
A replication competent avian-paramyxovirus minigenome comprising a first nucleic acid encoding a viral protein selected from the group consisting of NP, P and L and a second nucleic acid sequence corresponding to the 5′-terminal end, as set forth in SEQ ID NO:82, joined to a 3′-terminal end, as set forth in SEQ ID NO:80 or SEQ ID NO:81, of the genome of avian-paramyxovirus, wherein said second nucleic acid is a multiple of six nucleotides and capable of being transcribed, allowing replication of said second nucleic acid sequence.
A cDNA at least comprising:
- a first nucleic acid sequence as set forth in SEQ ID NO:82;
- a second nucleic acid sequence as set forth in SEQ ID NO:80 or SEQ ID NO:81;
- and,
- a third nucleic acid sequence, located between said first and second nucleic acid sequences, said third nucleic acid sequence having a length that is a multiple of six nucleotides and being transcribable.
Dependencies are inferred from the claim text.