Lonza Inc.

Lonza Ag

Morphosys Ag

Joachim Klein

Diethard Mattanovich

Brigitte Gasser

Roland Prielhofer

Michael Maurer

Jana Wenger

Johann Brass

Ralf Ostendorp

Armin Weidmann

Christoph Kiziak

A method of producing a protein of interest (POI) by culturing a recombinant eukaryotic cell line comprising an expression construct comprising a regulatable promoter and a nucleic acid molecule encoding a POI under the transcriptional control of said promoter, comprising the steps a) cultivating the cell line with a basal carbon source repressing the promoter, b) cultivating the cell line with a limited amount of a supplemental carbon source de-repressing the promoter to induce production of the POI at a transcription rate of at least 15% as compared to the native pGAP promoter, and c) producing and recovering the POI; and further an isolated regulatable promoter and a respective expression system.

Promoter variants for protein production

Methods of producing promoter variants

A method of producing a protein of interest (POI) by culturing a recombinant eukaryotic cell line comprising an expression construct comprising a regulatable promoter and a nucleic acid molecule encoding a POI under the transcriptional control of said promoter, comprising the steps a) cultivating the cell line with a basal carbon source repressing the promoter, b) cultivating the cell line with a limited amount of a supplemental carbon source de-repressing the promoter to induce production of the POI at a transcription rate or at least 15% as compared to the native pGAP promoter, and c) producing and recovering the POI; and further an isolated regulatable promoter and a respective expression system.

Regulatable promoter

Vectors expressible in a host that is the rhaBAD promoter region of the L-rhamnose operon operably linked to a transcriptional unit that is:

Rhamnose promoter expression system

New vectors expressible in a host comprising the promoter region of the melibiose operon operably linked to a transcriptional unit that includes nucleic acid sequence which is heterologous to the host. The expression of the nucleic acid sequence is controlled by the promoter region of the melibiose operon. The new vector can be used for the regulated heterologous expression of a nucleic acid sequence in a prokaryotic host. There is an isolated and purified nucleic acid sequence expressible in a host comprising the promoter region of the melibiose operon operably linked to a transcriptional unit that includes a nucleic acid sequence which is heterologous to the host. The expression of the nucleic acid sequence is controlled by the promoter region of the melibiose operon. A prokaryotic host is transformed with the vector or the isolated and purified nucleic acid sequence. There is a method for producing a polypeptide in a host using the vector.

Melibiose operon expression system

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Visp, Switzerland

Joachim Klein