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AssigneeHeineken Supply Chain B.V.
InventorsJean-Marc Georges Daran, Jacobus Thomas Pronk, Gabriele Romagnoli
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The invention relates to a nucleic acid molecule encoding a novel selection marker. Said marker is a guanidinobutyrase from Kluyveromyces lactis, which, when expressed in Saccharomyces, allows the growth of the yeast in the presence of guanidinobutyrate as the sole nitrogen source. Said marker can be used in a method for producing a microorganism having an altered genome. The invention further relates to a set of constructs, comprising a first construct comprising a recognition site for an endonuclease, a first region of homology with a target gene of a microorganism, and a first part of a nucleotide sequence encoding the selection marker, and a second construct comprising a second part of the nucleotide sequence encoding the selection marker, a second region of homology with the target gene of the microorganism, and a copy of the endonuclease recognition site. The invention further relates to methods for altering a target gene in a microorganism, to methods for producing a microorganism, and to microorganisms that are produced by the methods of the invention.
A method of culturing a microorganism of the genus Saccharomycetaceae in the presence of guanidinobutyrate as sole nitrogen source, comprising:
A microorganism of the genus Saccharomycetaceae that comprises a nucleotide sequence encoding a guanidinobutyrase.
A set of constructs, comprising a first construct comprising a first part of a nucleotide sequence encoding a guanidinobutyrase, and a second construct comprising a second part of the nucleotide sequence encoding the guanidinobutyrase, whereby a fragment of the first part of the nucleotide sequence overlaps with a fragment that is present in the second part of the nucleotide sequence, allowing recombination between the first and second part of the nucleotide sequence.
Dependencies are inferred from the claim text.