Monsanto Technology LLC

Calgene Corporation

Rutgers, the State University of New Jersey

Monsanto Company

Impetus Agriculture, Inc.

Jeffrey M Staub

Guangning Ye

Kevin E McBride

Larry A Gilbertson

Pal Maliga

David M Stalker

Yuechun Wan

Zora Svab Maliga

Vladimir Sidorov

Narender S Nehra

Debra L Broyles

James Kevin Roberts

William P Donovan

Edwards M Allen

Elysia K Krieger

Peter T Hajdukiewicz

Natalia Ivleva

Gregg Bogosian

David J Schaaf

Lori A Allison

Juan Luis Jurat-Fuentes

Helaine Carrer

Douglas A Russell

Julia P O'Neil

Oleg Zoubenko

Ivan Kanevski

Peter H J Hajdukiewicz

Constanze Hahnfeld

Heba Mohamed Yassen Abdelgaffar

Erik Jongedijk

Stefan Meldau

Debra Broyles

Affinity constructs and affinity molecules to direct insecticidal toxins to insect specific structures of target insects are presented herein. The affinity constructs comprise of at least one affinity molecule that is capable of recognizing, or capable of binding to, or binding to, or being directed to, or being designed to bind to an insect-specific structure in and/or on a target insect, and at least one other affinity molecule capable of binding to, or binding to, or being directed to, or being designed to bind to an insecticidal protein (toxin) wherein the at least two affinity molecules are operably coupled. Presented herein are also methods of making and using these affinity constructs and affinity molecules.

Affinity molecules and methods for their use

The disclosure provides nucleic acid constructs encoding novel chimeric peptides that are useful for nuclear and chloroplast expression of active nitrogenase components including dinitrogenase reductase and dinitrogenase, in plant cells. The disclosure thus provides nucleic acid constructs encoding the chimeric proteins, as well as plant expression constructs comprising the same for expression and/or targeting to the nucleus, plastids, or mitochondria of plant cells. The disclosure also provides methods of use for the novel chimeric peptides that function to provide active dinitrogenase reductase, dinitrogenase, and thus nitrogenase, expressed in plant cells, and plants comprising such nucleic acid constructs.

Compositions and methods for expression of nitrogenase in plant cells

A method is provided for transforming monocotyledonous plants to express DNA sequences of interest from plant cell plastids. The method allows the transformation of monocot plant tissue with heterologous DNA constructs. The invention also provides for monocot cells in which the plastids contain heterologous DNA constructs.

Plastid transformation of maize

A method is provided for transforming maize plants to express DNA sequences of interest from plant cell plastids. The method allows the transformation of maize plant tissue with heterologous DNA constructs by microprojectile bombardment of green light grown cultures and dark grown embryogenic cultures. The invention also provides for maize cells in which the plastids contain heterologous DNA constructs.

This invention discloses recombinant DNA constructs that transcribe to RNA having improved resistance to a plant RNase III enzyme, and methods for their use in making transgenic plant cells, plants, seeds, and products derived therefrom. Also provided are compositions and methods for imparting to a plant resistance to a pest or pathogen of the plant.

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Novel compositions and methods useful for genetic engineering of plant cells to provide a method of producing plastid transformed plants are provided in the instant invention. In particular, the present invention provides methods for obtaining plastid transformed plants on medium containing plastid lethal compounds.

Method for the transformation of plant cell plastids

Constructs and methods are provided for expressing peptides derived from eukaryotic organisms in plant plastids. Constructs have a promoter functional in a plant plastid, a DNA sequence encoding a peptide derived from an eukaryotic organism and a transcription termination region. Other elements include a selectable marker for selection of plant cells comprising a plastid expressing the marker and DNA regions of homology to the genome of the plastid and optionally a ribosome binding site joined to the promoter. By methods using such constructs high levels of eukaryotic peptides, such as mammalian proteins, are produced in a plant cell by growing plant cells under conditions whereby the DNA encoding sequences are expressed to produce eukaryotic peptide in said plastid.

Expression of eukaryotic peptides in plant plastids

Novel compositions and methods useful for genetic engineering of plant cells are provided. In particular, plastid constructs comprising recombining sites are provided. Such constructs find use in methods for site specific recombination in plant cell plastids. Plant cells and plants comprising the constructs described herein, as well as plant cells and plants produced by the methods of the present invention are of interest.

Methods and vectors for site-specific recombination in plant cell plastids

Constructs and methods are provided for expressing peptides derived from eukaryotic organisms in plant plastids. Constructs have a promoter functional in a plant plastid, a DNA sequence encoding a peptide derived from an eukaryotic organism and a transcription termination region. Other elements include a selectable marker for selection of plant cells comprising a plastid expressing the marker and DNA regions of homology to the genome of the plastid and optionally a ribosome binding site joined to the promoter. By methods using such constructs high levels of eukaryotic peptides, such as mammalian proteins, are produced in a plant cell by growing plant cells under conditions whereby the DNA encoding sequences re expressed to produce eukaryotic peptide in said plastid.

Provided are constructs and methods for expressing herbicide tolerance genes in plastids of plant cells. Constructs include the components of a promoter functional in a plant plastid, a DNA sequence which is capable of conferring tolerance in a plant cell to at least one herbicide compound when said DNA sequence is transcribed in plastids of said plant cell and a transcription termination region. Herbicide tolerance is produced by transforming plastids with the constructs of the invention and growing plant cells comprising the transformed plastids under conditions wherein the DNA sequence is transcribed and plant plastids and cells containing the plastids are rendered tolerant to applications of at least one herbicide compound.

Expression of herbicide tolerance genes in plant plastids

DNA constructs are provided for stable transformation of plastids of multicellular plants and expression of foreign proteins in plastids. The DNA constructs comprise a transforming DNA which is targeted to a pre-determined location on the plastid genome and inserted into the plastid genome by homologous recombination with targeting segments comprising DNA sequences homologous to the pre-determined region of the plastid genome. The transforming DNA contains a non-lethal selectable marker gene which confers a selectable phenotype on cells having plastids in which substantially all of the genomes therein contain the transforming DNA (i.e., homoplasmic cells or tissues). The transforming DNA further comprises at least one insertion site 4 for an additional DNA segment, such as a gene encoding a protein for improving a characteristic of the transformed plant. The non-lethal selectable marker gene is preferably provided as a chimeric gene by assembly from an expression cassette comprising 5&prime; and 3&prime; regulatory segments, preferably derived from plastid genes. A coding segment encoding the non-lethal selectable marker is inserted between the 5&prime; and 3&prime; regulatory segments to form the chimeric gene. The non-lethal selectable marker coding segment preferred in the present invention is the coding region of aadA from bacteria, which encodes aminoglycoside 3&Prime;-adenylyltransferase to confer spectinomycin and streptomycin resistance.

DNA constructs and methods for stably transforming plastids of multicellular plants and expressing recombinant proteins therein

Provided are methods for increasing the production of protein in a plant cell by transforming plastids of plant cells with a construct comprising a promoter functional in a plant plastid, a ribosome binding site, DNA sequence of interest and a transcription termination region, and growing plant cells comprising the transformed plastids under conditions wherein the DNA encoding sequence is transcribed in the plastid. Also provided are methods for increasing protein production by fusing a coding sequence to a gene of interest to a secondary protein for cleavage or targetting of the protein of interest within the plastid, whereby high levels of expression of protein is achieved.

Enhancer elements for increased translation in plant plastids

A composition for expression of a protein-encoding gene in a host cell is described, making use of an heterologous regulon. This composition provides a protein-encoding gene under control of a promoter heterologous to the host cell, and a gene for an RNA polymerase, preferably a single-subunit RNA polymerase that recognizes the promoter heterologous to the host cell. The gene for the RNA polymerase is under control of an inducible promoter recognized by the host cell. Also disclosed is a method for expressing the protein-encoding gene using the composition described.

Growing community of inventors

Wildwood, MO, United States of America

Jeffrey M Staub