Immuno Aktiengesellschaft

Baxter Aktiengesellschaft

Falko Gunter Falkner

Friedrich Dorner

Friedrich Scheiflinger

Michael Pfleiderer

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.

Direct molecular cloning of foreign genes into poxviruses and methods for the preparation of recombinant proteins

Methods for generating immune responses employing modified vaccinia of

An improved method is described to prepare recombinant fowlpox virus for the expression of proteins or for use as a vaccine. The new method uses for the insertion of foreign DNA an intergenic region which is located between the FPV thymidine kinase (tk)gene and the 3'-open reading frame. Said intergenic region is enlarged to comprise one or more unique restriction sites, thereby allowing insertion of foreign DNA in such a way that the FPV tk-gene remains intact and codes for the entire thymidine kinase. New strong poxvirus promoters are presented and new FPV host virus strains carrying a vaccinia virus thymidine kinase gene and an E. coli lacZ gene as a novel non-essential site. The novel fowlpox virus host strains allow the use of any insertion plasmid carrying vaccinia virus tk-gene flanking regions.

Recombinant fowlpox virus

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 most cell which packages the modified DNA molecule into infectious virions; and (III) recovering from the host cell virions comprised of the modified vital 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.

Growing community of inventors

Mannsdorf, Austria

Falko Gunter Falkner