Preparation of heterodimeric PDGF-AB using a bicistronic vector system in mammalian cells
AssigneesBeiersdorf AG, Gesellschaft fur Biotechnologische Forschung mbH
InventorsWolfram Eichner, Volker Achterberg, Albrecht Dorschner, Wolfgang Meyer-Ingold, Heiko Mielke, Wilhem Dirks, Manfred Wirth, Hansjorg Hauser
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Abstract
The recombinant production of PDGF-AB in mammalian cells is disclosed by means of a bicistronic vector system in which an IRES sequence is located between the first and second cistrons and in which the B chain coding gene is located in the first cistron. The disclosed expression unit allows the equimolor expression of the A and B polypeptide chains.
Claims12
A bicistronic expression unit for the recombinant preparation of heterodimeric PDGF-AB (Platelet Derived Growth Factor AB) in mammalian cells as host cells, characterized by the formula
- p-5'UTR-C.sub.1 -IRES-C.sub.2 -3'UTR-poly A,
- in which
- p is a transcriptional promoter,
- 5'UTR (5' untranslated region) is an untranslated nucleotide sequence,
- C.sub.1 is a cistron encoding the B chain of PDGF, a biologically active analog, or a biologically active fragment thereof,
- IRES (Internal Ribosomal Entry Site) is a nucleotide sequence of viral, cellular or synthetic origin that mediates an internal binding of the ribosomes,
- C.sub.2 is a cistron encoding the A chain of PDGF or a biologically active analog, or biologically active fragment thereof,
- 3'UTR (3' untranslated region) is an untranslated nucleotide sequence,
- and polyA is a polyadenylation signal,
- where the C.sub.1, IRES and C.sub.2 are connected to each other in an operative manner.
Dependencies are inferred from the claim text.