Selection in fungi
AssigneeNovozymes A/S of Krogshoejvej 36
InventorCarsten Lillelund Olsen
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Abstract
The present invention relates to methods for constructing a recombinant fungal host cell comprising one or more copies of a polynucleotide construct integrated in its genome, said method comprising transforming a fungal host cell with an integrative polynucleotide construct comprising a first polynucleotide encoding a selectable marker, wherein the first polynucleotide, a 5′ untranslated region thereof and/or a riboswitch operably linked therewith comprises a spliceosomal intron which has 5 nucleotides or less between its branch site and its acceptor site; and a second polynucleotide encoding a polypeptide of interest; as well as suitable polynucleotide constructs, resulting fungal host cells and methods of manufacture.
Claims28
A method for constructing a recombinant fungal host cell comprising one or more copies of a polynucleotide construct integrated in its genome, said method comprising:
- a) providing a fungal host cell transformed with an integrative polynucleotide construct, said construct comprising a first polynucleotide encoding a selectable marker, wherein the first polynucleotide, a 5′ untranslated region thereof and/or a riboswitch operably linked therewith comprises a spliceosomal intron which has 1 to 5 nucleotides between its branch site and its acceptor site;
- and a second polynucleotide encoding a polypeptide of interest;
- b) cultivating the transformed fungal host cell under conditions conducive for expressing the selectable marker;
- and
- (c) isolating a recombinant fungal host cell comprising one or more copies of the polynucleotide construct integrated in its genome.
Dependencies are inferred from the claim text.