Direct molecular cloning of foreign genes into poxviruses and methods for the preparation of recombinant proteins
US6265183A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Dec 19, 1994 |
| Grant date | Jul 24, 2001 |
| Priority date | — |
| Expiry date | Dec 19, 2014 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2740/16222
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
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 c…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.