Icosagen Cell Factory Ou

Kosagen Cell Factory Oü

Toomas Silla

Mart Ustav

Ene Ustav

Ingrid Tagen

Tiiu Mandel

Aare Abroi

Jelizaveta Geimanen

Kadri Janikson

Urve Toots

Andres Tover

Anne Kalling

Radi Tegova

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).

Viral expression plasmids for production of proteins, antibodies, enzymes, virus-like particles and for use in cell-based assays

This disclosure shows that the BPV-1 E2 protein-dependent minichromosome maintenance element (MME) comprised of E2 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). MME dependent plasmids are lost with the frequency of 6% per generation. Significantly long stable maintenance replication can also be provided without selection pressure. We also demonstrate that PyV core origin maintenance function/replication activation could be provided by Epstein-Barr virus Family of repeats and EBNA1 protein. The maintenance of the Polyomavirus core origin plasmid was characterized by 13% loss of the plasmid during one cell generation in the case of EBV FR harboring plasmids. Our data clearly indicate that maintenance functions from different viruses can provide segregation/partitioning function to different heterologous origins in variety of cells and can be used in expression of gene products.

Vectors, cell lines and their use in obtaining extended episomal maintenance replication of hybrid plasmids and expression of gene products

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Tartu, Estonia

Toomas Silla