Ura5 gene and methods for stable genetic integration in yeast
AssigneeGlycoFi, Inc.
InventorJuergen H. Nett
Life of this patent
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Abstract
A novel gene encoding P. pastoris orotate-phosphoribosyl transferase (URA5) is disclosed. Methods for producing and selecting yeast strains capable of stable genetic integration of heterologous sequences into the host genome are also provided.
Claims14
1-26. (canceled)
An isolated antibody or antigen-binding fragment or derivative thereof which binds selectively to the isolated polypeptide of claim 12, 14 or 15.
A method for the genetic integration of a heterologous nucleic acid sequence in a host cell comprising the step of disrupting a host gene encoding orotate-phosphoribosyl transferase by introduction of a disrupted, deleted or mutated nucleic acid sequence derived from a sequence selected from the group consisting of:
- (a) SEQ ID NO:2;
- (b) a nucleic acid sequence that is a degenerate variant of SEQ ID NO:2;
- (c) a nucleic acid sequence at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or at least 99.9% identical to SEQ ID NO:2;
- (d) a nucleic acid sequence that encodes a polypeptide having the amino acid sequence of SEQ ID NO:3;
- (e) a nucleic acid sequence that encodes a polypeptide at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or at least 99.9% identical to SEQ ID NO:3;
- (f) a nucleic acid sequence that hybridizes under stringent conditions to SEQ ID NO:2;
- and
- (g) a nucleic acid sequence comprising a fragment of any one of (a)-(f) that is at least 60 contiguous nucleotides in length.
A method for the genetic integration of a heterologous nucleic acid sequence in a host cell lacking orotate-phosphoribosyl transferase activity comprising the step of introducing a sequence of interest into the host cell in linkage with a sequence encoding orotate-phosphoribosyl transferase activity selected from the group consisting of:
- (a) SEQ ID NO:2;
- (b) a nucleic acid sequence that is a degenerate variant of SEQ ID NO:2;
- (c) a nucleic acid sequence at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or at least 99.9% identical to SEQ ID NO:2;
- (d) a nucleic acid sequence that encodes a polypeptide having the amino acid sequence of SEQ ID NO:3;
- (e) a nucleic acid sequence that encodes a polypeptide at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or at least 99.9% identical to SEQ ID NO:3;
- (f) a nucleic acid sequence that hybridizes under stringent conditions to SEQ ID NO:2;
- and
- (g) a nucleic acid sequence comprising a fragment of any one of (a)-(f) that is at least 60 contiguous nucleotides in length.
Dependencies are inferred from the claim text.