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AssigneeNovozymes Biotech, Inc.
InventorsDebbie S. Yaver, Daniel A. Bellini
Estimating the term…
The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a fungal host cell in a medium conducive for the production of the polypeptide; and (b) isolating the polypeptide from the cultivation medium; wherein the fungal host cell comprises a first nucleic acid sequence encoding the polypeptide in tandem with a second nucleic acid sequence comprising a crippled translational initiator sequence operably linked to a gene encoding a selectable marker, wherein the copy number of the first nucleic acid sequence has been increased by culturing the cell under conditions that select for multiple copies of the selectable marker. The present invention also relates to such fungal host cells and methods for obtaining such fungal host cells. The present invention further relates to nucleic acid constructs and vectors comprising a crippled translational initiator sequence operably linked to a gene encoding a selectable marker.
A method for producing a polypeptide, comprising:
A nucleic acid construct comprising in tandem a first nucleic acid sequence encoding a polypeptide and a second nucleic acid sequence comprising a crippled translational initiator sequence operably linked to a gene encoding a selectable marker in which the 3′ end of the crippled translational initiator sequence is immediately upstream of the initiator codon of the gene encoding the selectable marker, wherein the crippled translational initiator sequence comprises a T at the −3 position and a T at one or more of the −1, −2, and −4 positions.
A fungal host cell comprising a first nucleic acid sequence encoding a polypeptide in tandem with a second nucleic acid sequence comprising a crippled translational initiator sequence operably linked to a gene encoding a selectable marker in which the 3′ end of the crippled translational initiator sequence is immediately upstream of the initiator codon of the gene encoding the selectable marker, wherein the copy number of the first nucleic acid sequence has been increased by culturing the cell under conditions that select for multiple copies of the selectable marker, wherein the crippled translational initiator sequence comprises a T at the −3 position and a T at one or more of the −1, −2, and −4 positions.
Dependencies are inferred from the claim text.