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US 7,803,991 B2Granted
Universal chloroplast integration and expression vectors, transformed plants and products thereof
AssigneeAuburn University
InventorHenry Daniell
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AssigneeAuburn University
InventorHenry Daniell
Estimating the term…
The invention provides universal chloroplast integration and expression vectors which are competent to stably transform and integrate genes of interest into chloroplast genome of multiple species of plants. Transformed plants and their progeny are provided. Monocotyledonous and dicotyledonous plants are transformed which have never been transformed heretofore. Plants transformed with a synthetic gene express valuable biodegradable protein-based polymers (PBPs). Transformed plants produce high value molecules. Resistance is provided to agricultural crops against the major classes of chemical herbicides. Herbicide resistance is used as a lethal selectable marker for chloroplast transformation. The transformed plants are capable of expressing in addition to the targeted trait, a desirable, secondary non-targeted trait. Insect resistance is provided to transformed plants, both against insects that are susceptible to Bt toxins and against insects that have developed resistance to Bt toxins.
A stably transformed herbicide resistant target plant species, or the progeny thereof, which comprises a chloroplast stably transformed with a universal integration and expression vector competent for stable transformation of the chloroplast of different plant species, which vector comprises an expression cassette which encodes a heterologous protein of interest expressed by a heterologous DNA sequence in the chloroplast genome of the target plant species, which heterologous DNA confers tolerance to said herbicide, and flanking each side of the expression cassette, flanking DNA sequences derived from a transcriptionally active spacer region of a chloroplast genome, whereby stable integration of the heterologous DNA encoding the heterologous protein into a transcriptionally active spacer region of the chloroplast genome of the target plant was facilitated through homologous recombination of the flanking sequences with the homologous sequences in the target chloroplast genome.
A process for conferring herbicide resistance to a target plant species, which process comprises introducing into a plant cell of the target plant an integration and expression vector which vector comprises an expression cassette that comprises, operably joined, a heterologous DNA sequence coding for a protein of interest which confers resistance to a herbicide and control sequences positioned upstream from the 5′ and downstream from the 3′ ends of the coding sequence to provide expression of the coding sequence in the chloroplast of the target plant, a heterologous nucleotide sequence that confers a selectable phenotype other than tolerance to said herbicide, and flanking each side of the expression cassette, flanking sequences which are derived from a transcriptionally active spacer region of which are homologous to a transcriptionally active spacer sequence of the target chloroplast genome of the target plant, whereby stable integration of the heterologous coding sequence into a transcriptionally active spacer region of the chloroplast genome of the target plant is facilitated through homologous recombination of the flanking sequences with the homologous sequences in the target chloroplast genome;
A process of determining chloroplast transformation and expression of a target trait on the basis of a target plant species' acquisition of resistance to a selected herbicide due to the transformation of the plant species, which process comprises introducing into the plant a universal integration, and expression vector competent for stably transforming the chloroplast of different plant species, which vector comprises an expression cassette which comprises, operably joined, a heterologous DNA sequence coding for a protein conferring the desired target trait and control sequences positioned upstream from the 5′ and downstream from the 3′ ends of the coding sequence to provide expression of the coding sequence in the chloroplast of a target plant, a heterologous nucleotide sequence conferring herbicide resistance and flanking each side of the expression cassette, flanking DNA sequences which comprise each one a portion of the intergenic spacer 2 region between the tRNAIle and the tRNAAla genes of the chloroplast genome, which are homologous to a spacer sequence of the target chloroplast genome, which spacer sequence is conserved in the chloroplast genome of different plant species, whereby stable integration of the heterologous coding sequence into the chloroplast genome of the target plant is facilitated through homologous recombination of the flanking sequences with the homologous sequences in the target chloroplast genome;
Dependencies are inferred from the claim text.