Sembiosys Genetics, Inc.

Calgene Corporation

Her Majesty the Queen in Right of Canada, as Represented by the Minister

Commonwealth Scientific and Industrial Research Organisation

Uti Limited Partnership

Metabolix, Inc.

Semibiosys Genetics Inc.

Maurice M Moloney

Gijs Van Rooijen

Joseph Boothe

Janis Goll

Harm M Deckers

Sharon Radke

Joenel Alcantara

Youji Hu

Bipin Dalmia

Leonard Brent Selinger

Jin-Hao Liu

Kuo-Joan Cheng

Cecil Wallace Forsberg

Cory Lee Nykiforuk

Soheil Mahmoud

Surinder Pal Singh

Oliver P Peoples

Kristi D Snell

Nii Patterson

Hamid R Habibi

Sarita Chaudhary

Steven Szarka

Anthony B Schryvers

Wendy A Hutchins

Alexandra Reid

Richard Keon

Methods for the production of an apolipoprotein in plants are described. In one embodiment, the present invention provides a method for the expression of apolipoprotein in plants comprising:

Methods for the production of apolipoproteins in transgenic plants

The present invention relates to the use of a class of genes called oil body protein genes that have unique features. The discovery of these features allowed the invention of methods for the production of recombinant proteins wherein a protein of interest can be easily separated from other host cell components. The invention is further exemplified by methods for exploitation of the unique characteristics of the oil body proteins and oil body genes for expression of polypeptides of interest in many organisms, particularly plant seeds. Said polypeptides may include but are not limited to: seed storage proteins, enzymes, bioactive peptides, antibodies and the like. The invention can also be modified to recover recombinant polypeptides fused to oil body proteins from non-plant host cells. Additionally the invention provides a method of using recombinant proteins associated with seed oil bodies released during seed germination for expression of polypeptides that afford protection to seedlings from pathogens. Finally, the persistent association of oil body proteins with the oil body can be further utilized to develop a biological means to create novel immobilized enzymes useful for bioconversion of substrates.

Preparation of heterologous proteins on oil bodies

Novel methods for the expression of non-native genes in flax seeds and the seeds of other plant species are provided. The methods involve the use of seed-specific promoters obtained from flax. Additionally provided are novel flax seed-specific promoters, chimeric nucleic acid constructs comprising novel flax seed-specific promoters, transgenic plant cells, transgenic plants and transgenic plant seeds containing novel flax seed-specific promoters. The promoters and methods are useful, for example, for altering the seed oil and protein composition in flax seed or other plant seeds.

Flax seed specific promoters

The present invention provides novel emulsion formulations which comprise oil bodies. The invention also provides a method for preparing the emulsions and the use of the emulsions in a variety of products including food products, personal care products and pharmaceutical products. In a preferred embodiment the emulsions comprise thioredoxin and/or thioredoxin reductase.

Thioredoxin and thioredoxin reductase containing oil body based products

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.

Methods for the production of insulin in plants

An improved method for recovering recombinantly produced polypeptides is described. The method involves expressing the recombinant polypeptide as a fusion protein with a pro-peptide. The pro-peptide-polypeptide fusion protein can be cleaved and the recombinant polypeptide released under the appropriate conditions.

Method for cleavage of fusion proteins

Method for producing and cleaving a fusion proteins with an n-terminal chymosin pro-peptide

A method for the separation of a target molecule from a mixture is described. The method employs oil bodies and their associated proteins as affinity matrices for the selective, non-covalent binding of desired target molecules. The oil body proteins may be genetically fused to a ligand having specificity for the desired target molecule. Native oil body proteins can also be used in conjunction with an oil body protein specific ligand such as an antibody or an oil body binding protein. The method allows the separation and recovery of the desired target molecules due to the difference in densities between oil bodies and aqueous solutions.

Oil bodies and associated proteins as affinity matrices

Improved methods for the production of multimeric-protein-complexes, such as redox proteins and immunoglobulins, in association with oil bodies are described. The redox protein is enzymatically active when prepared in association with the oil bodies. Also provided are related nucleic acids, proteins, cells, plants, and compositions.

Methods for the production of multimeric immunoglobulins, and related compositions

The present invention provides a method of preparing epidermal growth factors in plants and transgenic plant seeds containing epidermal growth factors. The method provides an economical way to produce epidermal growth factors.

Expression of epidermal growth factor in plant seeds

The present invention provides novel adjuvants which comprise oil bodies. The invention also provides vaccine formulations comprising oil bodies and an antigen and methods for preparing the vaccines and the use of the vaccines to elicit an immune response.

Immunogenic formulations comprising oil bodies

The present invention relates to the use of a class of genes called oil body protein genes that have unique features. The discovery of these features allowed the invention of methods for the production of recombinant proteins wherein a protein of interest can be easily separated from other host cell components. The invention is further exemplified by methods for exploitation of the unique characteristics of the oil body proteins and oil body genes for expression of polypeptides of interest in many organisms, particularly plant seeds. Said polypeptides include thioredoxin and/or thioredoxin reductase. The invention can also be modified to recover recombinant polypeptides fused to oil body proteins from non-plant host cells.

Preparation of thioredoxin and thioredoxin reductase proteins on oil bodies

The present invention provides novel emulsion formulations which comprise oil bodies. The invention also provides a method for preparing the emulsions and the use of the emulsions in products for topical application to the skin. The products are very mild to the skin and may be easily formulated into a wide variety of personal care and dermatological products.

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Calgary, Canada

Maurice M Moloney