Yeast strains and methods for producing collagen
AssigneeModern Meadow, Inc.
InventorsLixin Dai, Julia Borden, Jeffrey Nelson, Kristin Ruebling-Jass
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Abstract
Strains of yeast genetically engineered to produce increased amounts of non-hydroxylated collagen or hydroxylated collagen are described. An all-in-one vector including the DNA necessary to produce collagen, promotors, and hydroxylating enzymes is also described. Methods for producing non-hydroxylated or hydroxylated collagen are also provided.
Claims33
A strain of yeast genetically engineered to produce non-hydroxylated collagen wherein the strain comprises a vector comprising an optimized DNA sequence encoding bovine collagen, wherein the DNA sequence is at least 90% identical to SEQ ID NO:
- 2.
A strain of yeast genetically engineered to produce hydroxylated bovine collagen wherein the strain comprises a first vector comprising an optimized DNA sequence encoding bovine collagen wherein the DNA sequence is at least 90% identical to SEQ ID NO:
- 2;
- and a second vector comprising a DNA sequence encoding P4HA1 and a DNA sequence encoding P4HB;
- and wherein the vectors have been inserted into the strain of yeast.
An all-in-one vector comprising:
- (i) an optimized DNA sequence encoding bovine collage, wherein the DNA sequence is at least 90% identical to SEQ ID NO:
- 2;
- (ii) a DNA sequence encoding hydroxylation enzymes comprising P4HA1, P4HB, and combinations thereof, including promoters and terminators;
- (iii) a DNA sequence for a selection marker, including a promoter and a terminator;
- (iv) a DNA sequence for origins of replication for yeast and bacteria;
- (v) DNA sequences with homology to a yeast genome for integration into the genome;
- and
- (vi) restriction sites at a position selected from the group consisting of 5′, 3 ′, within the above DNA sequences, and combinations thereof allowing for modular cloning.
Dependencies are inferred from the claim text.