Abstract: A plant produced vaccine for Porcine Epidemic Diarrhea Virus (PEDV) is provided where the Spike protein of the virus is expressed in a plant by introducing into a plant a construct comprising a promoter preferentially directing expression to seed of said plant, a nucleoic acid encoding the Spike protein and a nucleic acid targeting expression to the endoplasmic reticulum of the plant. The plant expresses the S1 polypeptide at levels of at least 10 mg/kg of seed of said plant. When orally administered to an animal, a protective response is observed including a serum antibody response.

Abstract: Increased antigenicity of a membrane-bound polypeptide produced from a plant is provided in a process in which extraction of the polypeptide or other compounds from the plant is such that phospholipids are associated with the polypeptide. Reducing fat by supercritical fluid extraction increases antigenicity of such plant-produced membrane-bound polypeptides. Methods and means of producing such membrane-bound polypeptides are provided. Methods to produce a protective response in animals are provided by administering to the animal the membrane-bound polypeptide. Binding of antibody specific to the membrane-bound polypeptide is increased. The process provides for increased preferred formation of the membrane-bound polypeptide. Stability of the membrane-bound polypeptide is increased when the plant material is defatted.

Abstract: Vaccines, methods of producing, and methods of using are provided in which a protective response to Valley Fever disease is produced when administered to an animal. The vaccine provides for expression of Coccidioides sp. Ag2 polypeptide in a plant or plant part, linked to a promoter preferentially directing expression to seed tissue of the plant or plant part. Further embodiments provide the polypeptide is targeted to the cell wall, vacuole or endoplasmic reticulum. The polypeptide may be fused to a dendritic cell targeting dendritic cell or a heat labile enterotoxin. Increased expression levels in the plant or plant part are obtained. The vaccine comprising the plant-produced Ag2 polypeptide may be a glucan chitin particle comprising the Ag2 polypeptide. The plant or plant materials in an embodiment may be orally administered.

Abstract: Methods of vaccination of animals are provided. In an embodiment, a paired administration of a primer vaccine provides for non-oral administration of a vaccine and an oral administration of the vaccine and can be followed by a paired administration of a booster vaccine of a non-oral administration and an oral administration. Embodiments provide the non-oral and oral administration are within three days of each other. The methods provide for improved protective response in an animal compared to administration of a primer non-oral administered vaccine followed by three booster non-oral administered vaccines. An adjuvant is provided that is a microcrystalline polysaccharide-based adjuvant derived from delta inulin.

Abstract: A plant produced vaccine for Porcine Epidemic Diarrhea Virus (PEDV) is provided where the Spike protein of the virus is expressed in a plant by introducing into a plant a construct comprising a promoter preferentially directing expression to seed of said plant, a nucleoic acid encoding the Spike protein and a nucleic acid targeting expression to the endoplasmic reticulum of the plant. The plant expresses the S1 polypeptide at levels of at least 10 mg/kg of seed of said plant. When orally administered to an animal, a protective response is observed including a serum antibody response.

Abstract: Methods of vaccination of animals are provided. In an embodiment, a paired administration of a primer vaccine provides for non-oral administration of a vaccine and an oral administration of the vaccine and can be followed by a paired administration of a booster vaccine of a non-oral administration and an oral administration. Embodiments provide the non-oral and oral administration are within three days of each other. The methods provide for improved protective response in an animal compared to administration of a primer non-oral administered vaccine followed by three booster non-oral administered vaccines. An adjuvant is provided that is a microcrystalline polysaccharide-based adjuvant derived from delta inulin.

Abstract: Vaccines, methods of producing, and methods of using are provided in which a protective response to Valley Fever disease is produced when administered to an animal. The vaccine provides for expression of Coccidioides sp. Ag2 polypeptide in a plant or plant part, linked to a promoter preferentially directing expression to seed tissue of the plant or plant part. Further embodiments provide the polypeptide is targeted to the cell wall, vacuole or endoplasmic reticulum. The polypeptide may be fused to a dendritic cell targeting dendritic cell or a heat labile enterotoxin. Increased expression levels in the plant or plant part are obtained. The vaccine comprising the plant-produced Ag2 polypeptide may be a glucan chitin particle comprising the Ag2 polypeptide. The plant or plant materials in an embodiment may be orally administered.

Abstract: A method of increasing expression of a product encoded by a nucleic acid molecule is provided where in one embodiment multiple plant transcription units comprising a promoter and the nucleic acid molecule are provided. In another embodiment multiple plant transcription units are provided where each promoter is different from the other. In an embodiment the multiple plant transcription units may comprise two, three, four or more plant transcription units. In another embodiment the promoter may be selected from an embryo promoter. Another embodiment provides the promoter may be a globulin promoter. A further embodiment provides the product encoded may be selected from hepatitis b, aprotinin, or a cellulase. Still further embodiments provide the product may be selected from the cellulases E1 and CBH1.

Abstract: Increased antigenicity of a membrane-bound polypeptide produced from a plant is provided in a process in which extraction of the polypeptide or other compounds from the plant is such that phospholipids are associated with the polypeptide. Reducing fat by supercritical fluid extraction increases antigenicity of such plant-produced membrane-bound polypeptides. Methods and means of producing such membrane-bound polypeptides are provided. Methods to produce a protective response in animals are provided by administering to the animal the membrane-bound polypeptide. Binding of antibody specific to the membrane-bound polypeptide is increased. The process provides for increased preferred formation of the membrane-bound polypeptide. Stability of the membrane-bound polypeptide is increased when the plant material is defatted.

Abstract: Increased antigenicity of a membrane-bound polypeptide produced from a plant is provided in a process in which extraction of the polypeptide or other compounds from the plant is such that phospholipids are associated with the polypeptide. Reducing fat by supercritical fluid extraction increases antigenicity of such plant-produced membrane-bound polypeptides. Methods and means of producing such membrane-bound polypeptides are provided. Methods to produce a protective response in animals are provided by administering to the animal the membrane-bound polypeptide. Binding of antibody specific to the membrane-bound polypeptide is increased. The process provides for increased preferred formation of the membrane-bound polypeptide. Stability of the membrane-bound polypeptide is increased when the plant material is defatted.

Abstract: An enhanced globulin-1 regulatory region is shown, a nucleotide sequence of which includes at least one additional copy of a region of the globulin-1 regulatory region which includes at least one transcriptional factor binding domain, combined with a transcription initiation site and translation start site. The promoter provides improved seed preferred, and particularly embryo preferred expression in plants. Methods of use are also shown in preferentially expressing a heterologous protein to the embryo tissue of a plant.

Abstract: Method and plants expressing increased levels of Butyrylcholinesterase (BChE) is described. The nucleic acid molecule encoding BChE is operably linked to a promoter preferentially expressing to the endosperm cells of the plant, another embodiment expression is targeted to the endoplasmic reticulum of plant cell(s), to the cell wall of the plant cell(s) or both.

Abstract: Increased antigenicity of a membrane-bound polypeptide produced from a plant is provided in a process in which extraction of the polypeptide or other compounds from the plant is such that phospholipids are associated with the polypeptide. Reducing fat by supercritical fluid extraction increases antigenicity of such plant-produced membrane-bound polypeptides. Methods and means of producing such membrane-bound polypeptides are provided. Methods to produce a protective response in animals are provided by administering to the animal the membrane-bound polypeptide. Binding of antibody specific to the membrane-bound polypeptide is increased. The process provides for increased preferred formation of the membrane-bound polypeptide. Stability of the membrane-bound polypeptide is increased when the plant material is defatted.

Abstract: Increased antigenicity of a membrane bound polypeptide produced from plants is provided by reducing fat content of the plant, plant part, or plant tissue producing the polypeptide. Methods and means of producing such plant material are provided. Methods to produce a protective immune response in animals are provided by administering to the animal the plant, plant part of plant tissue which has reduced fat content and which comprises the polypeptide or by administering to the animal an extracted polypeptide produced from such a plant.

Abstract: An enhanced globulin-1 regulatory region is shown, a nucleotide sequence of which includes at least one additional copy of a region of the globulin-1 regulatory region which includes at least one transcriptional factor binding domain, combined with a transcription initiation site and translation start site. The promoter provides improved seed preferred, and particularly embryo preferred expression in plants. Methods of use are also shown in preferentially expressing a heterologous protein to the embryo tissue of a plant.