Abstract: A method is provided for transforming undeveloped plastids in tobacco suspension culture to produce a transplastomic plant.

Abstract: 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.

Abstract: A method for improved plastid transformation efficiency via homologous recombination and nucleic acid sequences useful therefore is provided. Nucleic acid sequences comprising a 5 base pair recombination sequence motif or multiple direct repeats thereof that increase the frequency of integration of a selected transgene through plastid transformation by homologous recombination are provided.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.