CMV promoter and method for production of polypeptides
AssigneeHoffmann-La Roche Inc.
InventorsUlrich Goepfert, Benjamin Moritz
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Abstract
The current invention reports a promoter that has the nucleic acid sequence of SEQ ID NO: 02 or SEQ ID NO: 03 which is a human CMV major immediate-early (hCMV-MIE) promoter/enhancer with C to G point mutation at position −41 and/or −179 relative to the transcription start site. This new promoter is especially useful for the production of polypeptides at large scale as it shows reduced promoter silencing and improved polypeptide production.
Claims13
A promoter comprising the nucleic acid sequence of SEQ ID NO:
- 02 or SEQ ID NO:
- 03.
A nucleic acid molecule consisting of the nucleic acid sequence of SEQ ID NO:
- 02 or SEQ ID NO:
- 03, wherein the nucleic acid molecule is capable of having a promoter strength of at least 80% of the human CMV-major immediate-early promoter of SEQ ID NO:
- 01 when operably linked to the nucleic acid sequence of SEQ ID NO:
- 04.
A method of producing a polypeptide, comprising:
- a) transfecting a eukaryotic cell with a nucleic acid molecule comprising an expression cassette comprising a first nucleic acid sequence comprising the sequence of SEQ ID NO:
- 02 or SEQ ID NO:
- 03 operably linked to a second nucleic acid sequence encoding the polypeptide,
- b) selecting a cell transfected in step a),
- c) cultivating the selected cell of step b) in a cultivation medium, and
- d) recovering the polypeptide from the cell or the cultivation medium, thereby producing the polypeptide.
A promoter comprising the sequence of SEQ ID NO:
- 06.
Dependencies are inferred from the claim text.