C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/863 (2006.01) C07K 14/16 (2006.01) C07K 14/745 (2006.01) C07K 14/755 (2006.01) C12N 7/01 (2006.01) C12N 9/64 (2006.01) C12N 9/68 (2006.01) C12N 9/74 (2006.01) C12N 15/12 (2006.01) C12N 15/49 (2006.01) C12N 15/55 (2006.01) C12N 15/57 (2006.01) C12N 15/63 (2006.01) C12N 15/64 (2006.01) C12N 15/86 (2006.01) C12P 21/02 (2006.01)
Patent
CA 2076839
A method is disclosed for producing a modified eukaryotic cytoplasmic DNA virus by direct molecular cloning of a modified DNA molecule comprising a modified cytoplasmic DNA virus genome. The inventive method comprises the steps of (I) modifying under extracellular conditions a DNA molecule comprising a first cytoplasmic DNA virus genome to produce a modified DNA molecule comprising the modified cytoplasmic DNA virus genome; (II) introducing the modified DNA molecule into a first host cell which packages the modified DNA molecule into infectious virions; and (III) recovering from the host cell virions comprised of the modified viral genome. The host cell is infected with a helper virus which is expressed to package the modified viral genome into infectious virions. Examples of packaging a modified poxvirus genome by a helper poxvirus of the same or different genus are described. Also disclosed are novel poxvirus vectors for direct molecular cloning of open reading frames into a restriction enzyme cleavage site that is unique in the vector. In one model poxvirus vector, the open reading frame is transcribed by a promoter located in the vector DNA upstream of a multiple cloning site comprised of several unique cleavage sites.
Dorner Friedrich
Falkner Falko G.
Pfleiderer Michael
Scheiflinger Friedrich
Baxter Aktiengesellschaft
Baxter Healthcare S.a.
Fetherstonhaugh & Co.
Immuno Aktiengesellschaft
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