Novozymes A/s

Novo Nordisk A/s

Novozymes, Inc.

Novosymes A/s

Novid Nordisk

Michael Dolberg Rasmussen

Carsten Lillelund Olsen

Randy M Berka

Michael W Rey

Mads Eskelund Bjornvad

Peter Bjarke Olsen

Preethi Ramaiya

Allan Kent Nielsen

Jens Tønne Andersen

Jesper Vind

Helle Outtrup

Claus Von Der Osten

Henrik Bisgangrd-Frantzen

Pia Van Der Zee

Jon Martin Persson

Torben Vedel Borchert

Joel R Cherry

Poul Erik Pedersen

Per Lina Jorgensen

Henrik Bisgård-Frantzen

Peter Rahbek Stergaard

Stanislas Dusko Ehrlich

Peter Rahbek Østergaard

Anne Breuner

Gitte Bak Poulsen

Henrik Bisgard-Frantzen

Peter Rahbek Ýstergaard

Methods of constructing a cell comprising in its chromosome one or more copies of an open reading frame (ORF) or operon encoding at least one polypeptide of interest, each copy being under the transcriptional control of a heterologous promoter using a site specific recombinase and in vivo integration by recombination; resulting cells, and methods for producing a polypeptide of interest using the resulting cells.

Stable genomic integration of multiple polynucleotide copies

The present invention provides methods for directly transferring a recombinant polynucleotide of interest from the chromosome of a prokaryotic donor cell to the chromosome of a prokaryotic recipient cell of another species, wherein the polynucleotide of interest comprises at least one coding sequence of interest, preferably one gene of interest and a selectable marker, wherein said polynucleotide of interest in the chromosome of the donor cell is flanked on each side by a polynucleotide region derived from the chromosome of the recipient cell, and wherein the flanking polynucleotide regions are of sufficient size and sequence identity to allow double homologous recombination with the corresponding regions in the chromosome of the recipient cell.

Direct transfer of polynucleotides between genomes

A bacterial host cell is disclosed including at least two copies of an amplification unit in its genome, the amplification unit including: i) at least one copy of a gene of interest, and ii) an expressible conditionally essential gene, wherein the conditionally essential gene is either promoterless or transcribed from a heterologous promoter having an activity substantially lower than the endogenous promoter of the conditionally essential gene, and wherein the conditionally essential gene if not functional would render the cell auxotrophic for at least one specific substance or unable to utilize one or more specific sole carbon source; methods for producing a protein using the cell of the invention, and methods for constructing the cell of the invention.

Method for stable gene-amplification in a bacterial host cell

The present invention relates to cells producing at least one polypeptide of interest and expressing one or more recombinant nuclease encoding gene(s) thereby producing the nuclease(s), and methods for producing a polypeptide of interest essentially free from contaminating DNA, said method comprising the steps of:

DNase expression in recombinant host cells

A progeny cell derived from a parent cell, wherein the progeny cell comprises at least one gene encoding MrgA protein or a functional homologue thereof and/or a DNA segment operably linked with the encoding gene, wherein said gene and/or DNA segment is manipulated with respect to the parent cell; the progeny cell comprises two or more copies of a gene encoding MrgA protein or a functional homologue thereof; or the progeny cell is mutated with respect to the parent cell; whereby the progeny cell produces greater amounts of MrgA protein or a functional homologue thereof than the parent cell.

Cell with improved secretion mediated by MrgA protein or homologue

The present invention relates to cells producing at least one polypeptide of interest and expressing one or more recombinant nuclease encoding gene(s) thereby producing the nuclease(s), and methods for producing a polypeptide of interest essentially free from contaminating DNA, said method comprising the steps of: (a) cultivating a cell that produces at least one polypeptide of interest and expresses one or more recombinant nuclease encoding gene(s) thereby producing the nuclease(s); and (b) isolating the polypeptide of interest.

The present invention relates to an isolated polynucleotide of the complete chromosome of. The present invention also relates to isolated genes of the chromosome ofwhich encode biologically active substances and to nucleic acid constructs, vectors, and host cells comprising the genes as well as methods for producing biologically active substances encoded by the genes and to methods of using the isolated genes of the complete chromosome of

[object Object]

A mutated prokaryotic cell, which secretes higher amounts of at least one heterologous polypeptide of interest and which has a reduced expression-level of YusZ or YusX, or homologues thereof, when compared with an otherwise isogenic but non-mutated cell, and methods for constructing and using such a cell in the production of polypeptides.

Mutated prokaryotic cells with high secretion-levels

A bacterial host cell comprising at least two copies of an amplification unit in its genome, said amplification unit comprising: i) at least one copy of a gene of interest, and ii) an expressible conditionally essential gene, wherein the conditionally essential gene is either promoterless or transcribed from a heterologous promoter having an activity substantially lower than the endogenous promoter of said conditionally essential gene, and wherein the conditionally essential gene if not functional would render the cell auxotrophic for at least one specific substance or unable to utilize one or more specific sole carbon source; methods for producing a protein using the cell of the invention, and methods for constructing the cell of the invention.

The invention relates to a method for increasing the copy number of a chromosomally integrated expression cassette in a microbial strain without leaving antibiotic resistance markers behind in the strain, the necessary genetic constructs, and the strains resulting from the method of the invention.

Method for increasing gene copy number in a host cell and resulting host cell

Growing community of inventors

Vallensbaek, Denmark

Michael Dolberg Rasmussen