Methods for the production of insulin in plants
AssigneeSemBioSys Genetics Inc.
InventorsMaurice M. Moloney, Joseph Boothe, Richard Keon, Cory Nykiforuk, Gijs Van Rooijen
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Abstract
Commercial production of human insulin can be affected via transgenic expression in plant seeds. Thus, levels of insulin accumulation exceeding 0.1% of total cellular protein can be achieved recombinantly, through expression of the insulin with a single-chain antibody as a fusion partner. Production in seeds offers flexibility in storage and shipment of insulin as a raw material, and insulin retains its activity upon extraction from stored seed. Further, the amount of biomass subjected to extraction is limited, due to the relatively low water content of plant seeds.
Claims29
A method for the expression of insulin in plant seeds comprising:
- (a) providing a chimeric nucleic acid construct comprising in the 5′ to 3′ direction of transcription as operably linked components:
- (i) a first nucleic acid sequence that comprises a seed-preferred promoter;
- (ii) a second nucleic acid sequence encoding a signal peptide and an insulin polypeptide;
- and
- (iii) a third nucleic acid sequence that encodes a single chain antibody that has specificity for an oil body;
- (b) introducing the chimeric nucleic acid construct into a plant cell;
- (c) growing the plant cell into a mature plant capable of setting seed;
- and
- (d) obtaining substantially pure insulin from said seeds.
A plant capable of setting seed, wherein seeds of said plant comprise a chimeric nucleic acid sequence that comprises in the 5′ to 3′ direction of transcription:
- (a) a first nucleic acid sequence that comprises a seed-preferred promoter operatively linked to;
- (b) a second nucleic acid sequence encoding a signal peptide and an insulin polypeptide;
- and
- (c) a third nucleic acid sequence that encodes a single chain antibody that has specificity for an oil body.
A plant seed comprising a chimeric nucleic acid sequence that comprises in the 5′ to 3′ direction of transcription:
- (a) a first nucleic acid sequence that comprises a seed-preferred promoter that is operatively linked to;
- (b) a second nucleic acid sequence encoding a signal peptide and an insulin polypeptide;
- and
- (c) a third nucleic acid sequence that encodes a single chain antibody that has specificity for an oil body.
A polynucleotide comprised of (i) a first nucleic acid sequence that encodes insulin, and (ii) a second nucleic acid sequence that encodes a single chain antibody that has specificity for an oil body, and (iii) a third nucleic acid encoding a signal peptide operatively linked to (iv) a seed-preferred promoter.
A method for the commercial production of insulin, comprising
- (a) providing a plurality of seeds that comprise (i) a first polynucleotide coding for insulin operatively linked to (ii) a second polynucleotide that encodes a single chain antibody that has specificity for an oil body, and (iii) a third polynucleotide that encodes a signal peptide, wherein said seeds contain insulin;
- and
- (b) obtaining substantially pure insulin from said plurality.
A method for obtaining insulin-producing plants, comprising
- (a) providing a plurality of seeds that comprise (i) a first polynucleotide coding for insulin operatively linked to (ii) a second polynucleotide that encodes a single chain antibody that has specificity for an oil body, and (iii) a third polynucleotide that encodes a signal peptide, wherein said seeds contain insulin;
- and
- (b) using at least some seeds of said plurality to produce a population of plants, wherein plant of said population produce seeds that comprise insulin.
Dependencies are inferred from the claim text.