Use of homologous amdS genes as selectable markers
OwnerBart W. Swinkels, Gerardus C.M. Selten +3inventors
InventorsBart W. Swinkels, Gerardus C.M. Selten, Janna G. Bakhuis, Roelof A.L. Bovenberg, Adrianus W.H. Vollebregt
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Abstract
The present invention discloses novel amdS genes from fungi previously not known to contain and amdS gene, such as Aspergillus niger and Penicillium chrysogenum. The novel amdS genes can be used as homologous selectable marker genes in the transformation of these fungi. Alternatively, the cloned amdS genes can be used to inactivate the endogenous copy of the gene in order to reduce the background in transformation experiments.
Claims24
A DNA sequence encoding an acetamidase and characterized in that the DNA sequence is derived from an organism other than Aspergillus nidulans, Aspergillus oryzae and Saccharomyces cerevisiae.
A DNA sequence encoding an acetamidase which comprises an amino acid sequence which is characterized in that:
- a) its amino acid positional identity with one of the amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, and SEQ ID NO:5 is more than 30%, and
- b) its amino acid positional identity with each of the amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 is less than 100%.
A living cell selected from the group consisting of fungi from the Aspergillus niger group, the Aspergillus glaucus group, the Aspergillus terreus group, the Aspergillus restrictus group, the Aspergillus fumigatus group, the Aspergillus cervinus group, the Aspergillus ornatus group, the Aspergillus clavatus group, the Aspergillus versicolor group, the Aspergillus ustus group, the Aspergillus wentii group, the Aspergillus ochraceus group, the Aspergillus candidus group, the Aspergillus cremeus group, the Aspergillus sparsus group, Penicillium species, Trichoderma species, Mucor species, Rhizopus species, Phanerochaete species, Neurospora species, Humicola species, Claviceps species, Sordaria species, Ustilago species, Fusarium species, Schizophyllum species, Cephalosporium species, Acremonium species, edible fungi of which preferably Agaricus bisporus, Kluyveromyces species, Yarrowia species, Candida species, Hansenula species, Pichia species and Phaffia species, characterized by their ability to grow well on a culture medium containing acetamide as sole carbon and/or nitrogen source and wherein said ability is not caused by the expression of a heterologous acetamidase gene.
Dependencies are inferred from the claim text.