Recombinant modified vaccinia ankara (MVA) vaccinia virus containing restructured insertion sites
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsBernard Moss, Linda S. Wyatt, Patricia L. Earl
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Abstract
The present invention relates to recombinant modified vaccinia Ankara (MVA) virus containing restructured sites useful for the integration of heterologous nucleic acid sequences into an intergenic region (IGR) of the virus genome, where the IGR is located between two adjacent, essential open reading frames (ORFs) of the vaccinia virus genome, wherein the adjacent essential ORFs are non-adjacent in a parental MVA virus used to construct the recombinant MVA virus, and to related nucleic acid constructs useful for inserting heterologous DNA into the genome of a vaccinia virus, and further to the use of the disclosed viruses as a medicine or vaccine.
Claims20
An isolated nucleic acid construct comprising:
- (a) a first nucleic acid sequence derived from, or homologous to, a first essential ORF from a poxvirus genome;
- and
- (b) a second nucleic acid sequence derived from, or homologous to, a second essential ORF from a poxvirus genome;
- wherein the first and second essential poxvirus ORFs are separated by at least one non-essential ORF in the parental poxvirus genome, and wherein the first and second nucleic acid sequences are adjacent to each other in the isolated nucleic acid construct.
A method for producing a stable, recombinant poxvirus, the method comprising:
- (a) transfecting a cell with a nucleic acid construct comprising:
- i. a first nucleic acid sequence derived from, or homologous to, a first essential ORF from a poxvirus genome;
- and,
- ii. a second nucleic acid sequence derived from, or homologous to, a second essential ORF from a poxvirus genome;
- wherein the first and second essential poxvirus ORFs are separated by at least one non-essential ORF in the parental poxvirus genome, and wherein the first and second nucleic acid sequences are adjacent to each other in the isolated nucleic acid construct;
- (b) infecting the transfected cell with a poxvirus;
- (c) culturing the infected cell under conditions suitable to allow homologous recombination between the nucleic acid construct and the poxvirus genome.
Dependencies are inferred from the claim text.