Tyrosine prototrophy
AssigneePfizer Inc.
InventorsGregory Walter HILLER, Jeffrey Joseph MITCHELL, Bhanu Chandra MULUKUTLA, Pamela Mary PEGMAN
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Abstract
Provided herein is a tyrosine selection marker system, and uses thereof. In some embodiments, nucleic acid constructs, vectors, host cells and related compositions and methods for generating and selecting tyrosine prototroph cells are provided.
Claims32
A recombinant nucleic acid construct comprising a nucleotide sequence of interest, and one or both of:
- i) a phenylalanine hydroxylase (PAH) gene, and ii) a pterin-4-alpha-carbinolamine dehydratase (PCBD1) gene.
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A method of producing a host cell comprising an exogenous nucleotide sequence of interest, the method comprising:
- a) introducing into a host cell an exogenous nucleic acid construct comprising the nucleotide sequence of interest, wherein the exogenous nucleic acid construct further comprises:
- i) a PAH gene, and ii) a PCBD1 gene;
- b) culturing the host cell comprising the exogenous nucleic acid construct in a tyrosine-deficient medium, wherein the host cell comprising the exogenous nucleic acid construct proliferates more rapidly in the tyrosine-deficient medium than a corresponding otherwise identical host cell that lacks the exogenous nucleic acid construct.
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A method of producing a host cell comprising a first exogenous nucleotide sequence of interest and a second exogenous nucleotide sequence of interest, the method comprising:
- a) introducing into a host cell I) a first exogenous nucleic acid construct comprising i) the first exogenous nucleotide sequence of interest and ii) a PAH gene and II) a second exogenous nucleic acid construct comprising i) the second exogenous nucleotide sequence of interest and ii) a PCBD1 gene;
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- b) culturing the host cell comprising the first exogenous nucleic acid construct and the second exogenous nucleic acid construct in a tyrosine-deficient medium, wherein the host cell comprising the first exogenous nucleic acid construct and the second exogenous nucleic acid construct proliferates more rapidly in the tyrosine-deficient medium than a corresponding otherwise identical host cell that lacks the first exogenous nucleic acid construct and second exogenous nucleic acid construct.
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Dependencies are inferred from the claim text.