Back to results
US 8,895,808 B2Granted
Methods and vectors for producing transgenic plants
AssigneeMonsanto Technology LLC
InventorsLarry A. Gilbertson, Susan J. Johnson
Life of this patent
Estimating the term…
Abstract
Methods of, and compositions for, assembling one or more transcription units in a genome without a linked selectable marker or other unwanted transcription unit are provided. Also provided methods of, and compositions for, assembling one or more transcription units in a genome with a reduced frequency of vector backbone.
Claims18
A method of selecting for unlinked first and second DNA segments in a plant cell, comprising:
- a. transforming one or more plant cells with a nucleic acid molecule via Rhizobiaceae-mediated transformation, said nucleic acid molecule comprising:
- a first DNA segment comprising a first transcription unit located between a first left border region and a first right border region;
- a second DNA segment comprising a second transcription unit located between a second left border region and a second right border region, wherein said border regions are capable of enabling the integration of said segments into a plant genome;
- and
- a third transcription unit comprising a non-lethal negative selectable marker gene, wherein said non-lethal negative selectable marker gene is located between said first DNA segment and said second DNA segment, and wherein said left and right border regions are T-DNA border regions;
- b. growing said transformed one or more plant cells;
- and
- c. selecting from said transformed one or more plant cells a plant cell comprising said first DNA segment and said second DNA segment and lacking said non-lethal negative selectable marker gene.
5 more elements
A method of selecting for unlinked first and second DNA segments in a plant cell, comprising:
- a. transforming one or more plant cells with a nucleic acid molecule via Rhizobiaceae-mediated transformation, said nucleic acid molecule comprising:
- a first DNA segment comprising a first transcription unit located between a first T-DNA left border region and a first T-DNA right border region;
- a second DNA segment comprising a second transcription unit located between a second T-DNA left border region and a second T-DNA right border region, wherein said T-DNA border regions are capable of enabling the integration of said segments into a plant genome;
- and
- a third transcription unit comprising a non-lethal negative selectable marker gene, wherein said non-lethal negative selectable marker gene is located between said first DNA segment and said second DNA segment;
- b. growing said transformed one or more plant cells;
- c. selecting from said transformed one or more plant cells a plant cell comprising said first DNA segment and said second DNA segment and lacking said non-lethal negative selectable marker gene;
- and
- d. determining the linkage status of said first DNA segment and said second DNA segment in said selected plant cell and selecting for desired plant cells with unlinked said first DNA segment and said second DNA segment.
6 more elements
A recombinant nucleic acid molecule comprising:
- a first DNA segment comprising a first transcription unit located between a first left border region and a first right border region;
- a second DNA segment comprising a second transcription unit located between a second left border region and a second right border region, wherein said border regions are capable of enabling the integration of said segments into a plant genome, and wherein said left and right border regions are T-DNA border regions;
- and
- a third transcription unit comprising a non-lethal negative selectable marker gene, wherein said non-lethal negative selectable marker gene is located between said first DNA segment and said second DNA segment.
1 more element
Dependencies are inferred from the claim text.