Viral expression plasmids for production of proteins, antibodies, enzymes, virus-like particles and for use in cell-based assays
AssigneeIcosagen Cell Factory OÜ
InventorsToomas Silla, Ingrid Tagen, Anne Kalling, Radi Tegova, Mart Ustav, Tiiu Mandel, Urve Toots, Andres Tover, Aare Abroi, Ene Ustav, Jelizaveta Geimanen, Kadri Janikson
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Abstract
This disclosure shows that the EBV FR-element comprised of EBNA1 multimeric binding sites can provide the stable maintenance replication function to the mouse polyomavirus (PyV) core origin plasmids in the presence of BPV-1 E2 protein and PyV large T-antigen (LT).
Claims20
An expression system to provide extended episomal replication of hybrid plasmid in eukaryotic cell lines, said system comprising:
- a. a vector having a polyoma virus core origin consisting of SEQ ID NO:2, an FR element of EBV, a selection marker, and a gene of interest operably linked within an expression cassette expressing the gene in a eukaryotic cell;
- and
- b. a compatible mammalian cell line, where the vector is expressed, said cell line constitutively expressing EBV EBNA 1 protein and PyV LT protein either in presence or in absence of selective pressure.
An expression plasmid for use in mammalian cell expression system, said vector comprising:
- a. a polyoma virus core origin consisting of SEQ ID NO:2;
- b. an FR element of Eppstein Barr Virus (EBV);
- c. a selection marker;
- and
- d. a gene of interest in an expression cassette expressing in a eukaryotic cell.
Dependencies are inferred from the claim text.