High molecular weight collagen-like protein polymers
AssigneeProtein Polymer
InventorsJoseph Cappello, Franco A. Ferrari
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Abstract
Collagen-like polymers having repetitive triads are produced having reduced proline content, where glycine is the initial amino acid and the subsequent amino acids are varied, while retaining at least a minimum percentage of prolines. The resulting polymers have collagen-like properties, but are capable of being produced in unicellular microorganisms at high molecular weights and in high efficiency. The polymers while retaining collagen-like characteristics, include various novel sequences which impart new characteristics, finding wide use in photographic, medical, structural and fiber applications.
Claims21
A collagen like polymer of at least about 30 kD, characterized by:
- being expressed in a unicellular organism from a construct prepared in vitro;
- comprising at least 80 weight % of triads having glycine as the first amino acid and at least 40 number % of said triads comprising at least one proline, wherein the total proline content of said triads is less than 45 number %;
- and
- comprising at least two repetitive units of (glyXO).sub.n, wherein X and O symbolize individual amino acids, X and O are the same or different amino acids in each triad and n is at least 4;
- wherein at least two triads are selected from the group consisting of the triads gly-ala-pro, gly-pro-ala, gly-pro-pro, gly-ala-ser, gly-pro-gly, gly-pro-ser, gly-ala-gln, gly-ser-pro, gly-ser-gln, gly-leu-gln, gly-pro-arg, gly-pro-lys, gly-ala-lys, gly-ala-arg, gly-glu-arg, gly-asp-arg, gly-glu-pro, gly-asp-ala, gly-ala-his and gly-glu-ala.
A collagen like polymer of at least about 30 kD, characterized by:
- being expressed in a unicellular organism from a construct prepared in vitro;
- comprising at least 80 weight % of triads having glycine as the first amino acid and at least 40 number % of said triads comprising at least one proline and the proline total content of said triads is less than 45 number %;
- and
- comprising at least two repetitive units of the formula .alpha..sub.2 .beta..sub.n .alpha..sub.2, wherein .alpha. has three triads characterized as having glycine as the first amino acid and at least two of the triads having a proline;
- .beta. has from three to nine triads, wherein at least 30% of the triads have proline;
- and n is at least 6 and not more than about 36.
A unicellular microorganism comprising a DNA sequence encoding a collagen like polymer of at least about 30 kD and fewer than 45 number % proline said DNA sequence being characterized by:
- being expressed in said unicellular organism from a construct prepared in vitro;
- and
- comprising at least 80 weight % of triads having glycine as the first amino acid and at least 40 number % of said triads comprising at least one proline and the total proline content of said triads is less than 45 number %;
- and comprising at least two repetitive units of (glyXO).sub.n, wherein X and O are amino acids and are the same or different in each triad and n is at least 4.
A DNA sequence comprising a gene encoding a collagen like polymer of at least about 30 kD, characterized by:
- capable of being expressed in a unicellular organism from a construct prepared in vitro;
- comprising at least 80 weight % of triads having glycine as the first amino acid;
- and
- comprising at least two repetitive units of (glyXO).sub.n, wherein X and O are the same or different in each triad and n is at least 6 where X and O are selected to have from 15 to 45 number percent proline in said polymer;
- a transcriptional initiation regulatory region functional in a unicellular microorganism 5' of said gene;
- and
- a transcriptional termination regulatory region functional in a unicellular microorganism 3' of said gene;
- wherein said gene is under the transcriptional regulation of said regulatory regions.
A formed object comprising a collagen like polymer of at least about 30 kD, characterized by:
- being expressed in a unicellular organism from a construct prepared in vitro;
- comprising at least 80 weight % of triads having glycine as the first amino acid;
- and
- comprising at least two repetitive units of (glyXO).sub.n, wherein X and O are the same or different in each triad and n is at least 4 wherein X and O are selected to have from about 10 to 45 number % proline in said polymer.
Dependencies are inferred from the claim text.