RASAMSONIA TRANSFORMANTS
A method for carrying out recombination at a target locus in a Rasamsonia cell, which method comprises: - providing two or more nucleic acids which, when taken together, comprise: (a) sequences capable of homologous recombination with sequences flanking the target locus; (b) two or more site-specific recombination sites; (c) a sequence encoding a recombinase which recognizes the site-specific recombination sites; and (d) a sequence encoding a marker, wherein the two or more nucleic acids are capable of homologous recombination with each other so as to give rise to a single nucleic acid, and wherein at least two of the two or more nucleic acids each comprise a sequence encoding a non- functional portion of the marker; and - recombining in a Rasamsonia cell the said two or more nucleic acids with each other and with the sequences flanking the target locus so that a contiguous nucleic acid sequence encoding a functional marker and the sequence encoding the recombinase are inserted at the target locus, said marker-encoding and/or recombinase-encoding sequence being flanked by at least two site-specific recombination sites and the said site-specific recombination sites being flanked by the sequences capable of homologous recombination with sequences flanking the target locus, thereby to carry out recombination at a target locus in a Rasamsonia cell. The method may be used to generate marker free Rasamsonia cells.