Phillips Petroleum Company

Keck Graduate Institute

Alderbio Holdings LLC

Alder Biopharmaceuticals, Inc.

Research Corporation Technologies, Inc.

Biogrammatics, Inc.

H. Lundbeck A/s

Henkel Research Corporation

Keck Graduate Institute of Applied Life Sciences

James M Cregg

Ilya I Tolstorukov

John A Latham

Ilya I Tolstrorukov

Michael Miller Harpold

John Latham

Mark Litton

Randall Schatzman

Knut R Madden

Thomas G Chappell

George T Sperl

David W Stroman

Martin Anthony Gleeson

Juerg F Tschopp

Richard G Buckholz

Stephen Picataggio

Mary E Digan

Randal Schatzman

Lisa Haas

Randall Schatzman′

Kevin J Barringer

Iiya I Tolstorukov

Randall Schatzmann

In accordance with the invention, isolated nucleic acids, expression methods, host cells, expression vectors, and DNA constructs for producing proteins, and proteins produced using the expression methods are described. More particularly, nucleic acids isolated fromwherein the nucleic acids have promoter activity are described. The invention also relates to expression methods, host cells, expression vectors, and DNA constructs, for using thepromoters to produce proteins and polypeptides, and to the proteins and polypeptides produced using the expression methods.

Yeast promotors for protein expression

Isolated nucleic acids, expression methods, host cells, expression vectors, and DNA constructs for producing proteins, and proteins produced using the expression methods are disclosed. More specifically, nucleic acids isolated frompastons having promoter activity and expression methods, host cells, expression vectors, and DNA constructs of using thepastons promoters to produce different proteins and polypeptides are disclosed.

Methods are provided for the synthesis and secretion of recombinant hetero-multimeric proteins in mating competent yeast. A first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

Methods of synthesizing heteromultimeric polypeptides in yeast using a haploid mating strategy

Yeast promoters for protein expression

In accordance with the invention, isolated nucleic acids, expression methods, host cells, expression vectors, and DNA constructs for producing proteins, and proteins produced using the expression methods are described. More particularly, nucleic acids isolated fromwherein the nucleic acids have promoter activity are described. The invention also relates to expression methods, host cells, expression vectors, and DNA constructs, for using thepromoters to produce proteins, and to the proteins produced using the expression methods.

Yeast promoters from

Methods are provided for the synthesis and secretion of recombinant proteins preferably large mammalian proteins or hetero-multimeric proteins at high levels and for prolonged time in polyploid, preferably diploid yeast. These methods use various mating competent yeast, including. In a preferred embodiment, a first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

Novel promoters which are derived fromwhich are inducible or repressible under specific growth conditions are provided. These promoters are useful for regulating the expression of a desired structural gene, e.g., a mammalian polypeptide. Particularly preferred is the use of these novel promoters to regulate gene expression in polyploidal yeast such as diploidalproduced by mating or spheroplast fusion.

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Methods are provided for the synthesis and secretion of recombinant proteins preferably large mammalian proteins or hetero-multimeric proteins at high levels and for prolonged time in polyploid, preferably diploid yeast. These methods use various mating competent yeast, including Pichia. In a preferred embodiment, a first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

The present invention provides formaldehyde dehydrogenase genes (FLD) from methylotrophic yeasts. The FLD structural genes confer resistance to formaldehyde and are therefore useful as a selectable marker in methylotrophic yeasts. The FLD promoter sequences are strongly and independently induced by either methanol as sole carbon source (with ammonium sulfate as nitrogen source) or methylamine as sole nitrogen source (with glucose as carbon source). Induction under either methanol, methylamine or both provides levels of heterologous gene expression comparable to those obtained with the commonly used alcohol oxidase I gene promoter (P). The FLD promoter of(P) is an attractive alternative to Pfor expression of foreign genes in, allowing regulation by carbon (methanol) or nitrogen (methylamine) source within the same expression strain. Yeast strains, expression cassettes, expression vectors, and host cells-comprising an FLD gene promoter and 3' termination sequence are also provided.

Formaldehyde dehydrogenase genes from methylotrophic yeasts

Growing community of inventors

Claremont, CA, United States of America

James M Cregg