Abstract: Compositions and methods for regulating expression of nucleotide sequences of interest in a plant are provided. Compositions include novel nucleic acid molecules, and variants and fragments thereof, for expression control elements isolated from the Lemnaceae ubiquitin, r-histone and chitinase genes. A method for expressing a nucleotide sequence of interest in a plant using the expression control elements disclosed herein is further provided. The method includes introducing into a plant or plant cell or nodule an expression construct comprising an expression control element of the present invention operably linked to a nucleotide sequence of interest. In particular, the compositions and methods find use in enhancing expression of nucleotide sequences of interest in duckweed. Also provided is a novel Lemnaceae signal peptide-encoding sequence and the signal peptide encoded thereby.

Abstract: Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.

Abstract: Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. The methods comprise introducing into the plant a recombinant construct that provides for the inhibition of expression of ?1,3-fucosyltransferase (FucT) and ?1,2-xylosyltransferase (XylT) in a plant. Use of these constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a “humanized” N-glycosylation pattern without impacting plant growth and development. Stably transformed higher plants having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the G0 glycoform, are also provided.

Abstract: Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. The methods comprise introducing into the plant a recombinant construct that provides for the inhibition of expression of ?1,3-fucosyltransferase (FucT) and ?1,2-xylosyltransferase (XylT) in a plant. Use of these constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a “humanized” N-glycosylation pattern without impacting plant growth and development. Stably transformed higher plants having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the G0 glycoform, are also provided.

Abstract: Compositions and methods for regulating expression of nucleotide sequences of interest in a plant are provided. Compositions include novel nucleic acid molecules, and variants and fragments thereof, for expression control elements isolated from the Lemnaceae ubiquitin, r-histone and chitinase genes. A method for expressing a nucleotide sequence of interest in a plant using the expression control elements disclosed herein is further provided. The method includes introducing into a plant or plant cell or nodule an expression construct comprising an expression control element of the present invention operably linked to a nucleotide sequence of interest. In particular, the compositions and methods find use in enhancing expression of nucleotide sequences of interest in duckweed. Also provided is a novel Lemnaceae signal peptide-encoding sequence and the signal peptide encoded thereby.

Abstract: The present invention provides biologically active variants of human ?-2b-interferon. The variants contain carboxy terminus truncations when compared with the amino acid sequence of full-length human ?-2b-interferon. It is the novel finding of the present invention that these truncated variants have the biological activity of full-length human ?-2b-interferon. The invention encompasses these biologically active variant ?-interferons, as well as polynucleotides encoding these interferons. Expression cassettes comprising these polynucleotides and host cells comprising the expression cassettes are also provided. The invention also provides compositions comprising variant ?-interferon polypeptides and a pharmaceutically acceptable carrier.

Abstract: A bioreactor assembly of the present invention is provided for holding a media and supporting growth of a plurality of plants. The assembly includes a light source and a container having a light transmissive wall structure and defining a reservoir. A major axis of the reservoir is substantially horizontal allowing the reservoir to be filled with media to a partial level and to define a relatively large surface area for support of the plants. The assembly may also include clamps to secure and seal separate wall structure portions of the container together, and end caps to the wall portions, to define an aseptic environment. As another option, the clamps may define openings therethrough that allow passage of various devices for measuring and controlling bioreactor function such as a gas supply nozzle, a gas exit nozzle, an air temperature probe, a pH probe, a sampling drain, a gas composition probe and a media temperature probe.

Abstract: A bioreactor assembly of the present invention for holding a media and supporting growth of a plurality of plants. The assembly includes a light source and a container having a light transmissive wall structure and defining a reservoir. A major axis of the reservoir is substantially horizontal allowing the reservoir to be filled with media to a partial level and to define a relatively large surface area for support of the plants. The assembly may also include clamps to secure and seal separate wall structure portions of the container together, and end caps to the wall portions, to define an aseptic environment. As another option, the clamps may define openings therethrough that allow passage of various devices for measuring and controlling bioreactor function such as a gas supply nozzle, a gas exit nozzle, an air temperature probe, a pH probe, a sampling drain, a gas composition probe and a media temperature probe.

Abstract: A bioreactor assembly of the present invention for holding a media and supporting growth of a plurality of plants. The assembly includes a light source and a container having a light transmissive wall structure and defining a reservoir. A major axis of the reservoir is substantially horizontal allowing the reservoir to be filled with media to a partial level and to define a relatively large surface area for support of the plants. The assembly may also include clamps to secure and seal separate wall structure portions of the container together, and end caps to the wall portions, to define an aseptic environment. As another option, the clamps may define openings therethrough that allow passage of various devices for measuring and controlling bioreactor function such as a gas supply nozzle, a gas exit nozzle, an air temperature probe, a pH probe, a sampling drain, a gas composition probe and a media temperature probe.

Abstract: An assembly for promoting the growth of plant tissues that includes a plurality of plates each defining an array of wells wherein each of the wells contains a tissue sample. Support for the plates is provided by a rack having a plurality of vertically stacked shelves that may include one or more register depressions that urge the plates into predetermined positions. Light for the tissue samples is provided by a plurality of light-emitting diode arrays each mounted on a circuit board. Each circuit board is supported by a respective card edge connector of the rack so that the light-emitting diodes are in proximity to the plates supported on one of the shelves therebelow. Preferably, the light-emitting diode array corresponds to the well array supported in the registered position on the shelf therebelow so that each light-emitting diode is centered above a respective one of the wells.

Abstract: Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.

Abstract: The present invention provides methods and compositions for the production of recombinant plasminogen, microplasminogen, and fragments thereof in a duckweed expression system. It is the novel finding of the present invention that a duckweed expression system may be used to produce high levels of plasminogen and microplasminogen. The duckweed-produced plasminogen and microplasminogen can be activated to produce a polypeptide having protease activity. Thus, the invention encompasses methods for the expression of plasminogen, microplasminogen, and fragments thereof in duckweed, duckweed plants that are transformed with expression cassettes for the expression of plasminogen, microplasminogen, and fragments thereof, and nucleic acids comprising nucleotide sequences encoding plasminogen, microplasminogen, and fragments thereof, where these nucleotide sequences are modified to enhance their expression in duckweed.

Abstract: Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.

Abstract: The present invention provides methods and compositions for the transformation of duckweed plastids. The compositions and methods of the invention are useful in increasing the recombinant protein production capacity of the duckweed expression system. The compositions of the invention include transformed duckweed plastids and transplastomic duckweed cells and plants, as well as nucleic acid constructs useful for transforming duckweed plastids. The invention also provides methods for introducing one or more heterologous nucleotide sequences into a duckweed plastome.

Abstract: Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium. Methods of producing recombinant monoclonal antibodies having effector function, and antibodies produced by these methods are provided.

Abstract: A bioreactor assembly of the present invention for holding a media and supporting growth of a plurality of plants. The assembly includes a light source and a container having a light transmissive wall structure and defining a reservoir. A major axis of the reservoir is substantially horizontal allowing the reservoir to be filled with media to a partial level and to define a relatively large surface area for support of the plants. The assembly may also include clamps to secure and seal separate wall structure portions of the container together, and end caps to the wall portions, to define an aseptic environment. As another option, the clamps may define openings therethrough that allow passage of various devices for measuring and controlling bioreactor function such as a gas supply nozzle, a gas exit nozzle, an air temperature probe, a pH probe, a sampling drain, a gas composition probe and a media temperature probe.

Abstract: Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.

Abstract: An assembly for promoting the growth of plant tissues that includes a plurality of plates each defining an array of wells wherein each of the wells contains a tissue sample. Support for the plates is provided by a rack having a plurality of vertically stacked shelves that may include one or more register depressions that urge the plates into predetermined positions. Light for the tissue samples is provided by a plurality of light-emitting diode arrays each mounted on a circuit board. Each circuit board is supported by a respective card edge connector of the rack so that the light-emitting diodes are in proximity to the plates supported on one of the shelves therebelow. Preferably, the light-emitting diode array corresponds to the well array supported in the registered position on the shelf therebelow so that each light-emitting diode is centered above a respective one of the wells.

Abstract: An assembly for promoting the growth of plant tissues that includes a plurality of plates each defining an array of wells wherein each of the wells contains a tissue sample. Support for the plates is provided by a rack having a plurality of vertically stacked shelves that may include one or more register depressions that urge the plates into predetermined positions. Light for the tissue samples is provided by a plurality of light-emitting diode arrays each mounted on a circuit board. Each circuit board is supported by a respective card edge connector of the rack so that the light-emitting diodes are in proximity to the plates supported on one of the shelves therebelow. Preferably, the light-emitting diode array corresponds to the well array supported in the registered position on the shelf therebelow so that each light-emitting diode is centered above a respective one of the wells.

Abstract: Methods for high-throughput screening in duckweed are disclosed. In one aspect, these methods are used to identify nucleotide sequences encoding polypeptides of interest. In another aspect, these methods are used to identify nucleotide sequences that modulate the expression of a target nucleotide sequence. The methods combine the predictive benefits of screening in whole plants with the speed and efficiency of a high throughput system.