Infectious clone of human parvovirus B19 and methods
AssigneeGovernment of the United States, as represented by the Secretary, Dept. of Health and Human Services
InventorsKevin Brown, Ning Zhi, Peter Tijssen, Zoltan Zadori
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Abstract
The invention relates to infectious clones of parvovirus B19, methods of cloning infectious B19 clones, and methods of cloning viral genomes that have secondary DNA structures that are unstable in bacterial cells. A B19 infectious clone and methods of producing B19 infectious clones are useful for producing infectious virus. Infectious virus is useful for identifying and developing therapeutically effective compositions for treatment and/or prevention of human parvovirus B19 infections.
Claims45
A method for cloning a parvovirus B19 viral genome comprising:
- (a) introducing a vector comprising all or a portion of a parvovirus B19 genome into a prokaryotic cell that is deficient in at least one recombinase enzyme;
- (b) incubating the cells at about 25° C. to 35° C.;
- and
- (c) recovering the vector from the prokaryotic cells.
The cells are incubated for a time sufficient to allow amplification of the vector.
A method for producing an infectious virus of parvovirus B19, comprising:
- (a) introducing a vector comprising an infectious clone of parvovirus B19 into a population of cells, wherein the vector is present in at least about 15% of the cells;
- and
- (b) incubating the cells under conditions to allow for viral replication.
An isolated infectious parvovirus B19 clone comprising all or a portion of a parvovirus B19 viral genome and a replicable vector.
A method of identifying an infectious clone comprises:
- introducing a vector comprising all or a portion of a viral genome into a eukaryotic cell;
- incubating the cell for a sufficient time to produce infectious virus;
- and detecting production of infectious virus.
Dependencies are inferred from the claim text.