Selectable marker genes
The subject invention relates to a novel gene referred to herein as DSM-2. This gene was identified in Sterptomyces coelicolor A3. The DSM-2 protein is distantly related to PAT and BAR. The subject invention also provides plant-optimized genes encoding DSM-2 proteins, DSM-2 can be used as a transgenic trait to impart tolerance in plants and plant cells to the herbicides glufosinate and bialaphos. One preferred use of the subject genes are as selectable markers. The use of this gene as a selectable marker in a bacterial system can increase efficiency for plant transformations. Use of DSM-2 as the sole selection marker eliminates the need for an additional medicinal antibiotic marker (such as ampicillin resistance) during cloning. Various other uses are also possible according to the subject invention.
1. A phosphinothricin-resistant transgenic plant cell comprising between 1 and 3 copies of the polynucleotide of SEQ ID NO:3 integrated in the genomic DNA of the plant cell, wherein the plant cell is resistant to glufosinate when applied at a rate of 560 g ai/ha.
2. A method of selecting for a transgenic plant cell, the method comprising:
transforming a plurality of plant cells with a low copy number vector comprising a polynucleotide operably linked to a promoter operable in the plant cells, wherein the polynucleotide is SEQ ID NO:3, thereby producing transgenic plant cells comprising a low copy number of the polynucleotide; and
growing the plurality of plant cells in a concentration of bialaphos or glufosinate that permits cells that express the polynucleotide to grow, while killing or inhibiting the growth of cells that do not express the polynucleotide,
thereby selecting a transgenic plant cell that is resistant to glufosinate when applied at a rate of 560 g ai/ha.
3. The method of claim 2 , wherein the method further comprises regenerating a plant from the transgenic plant cell.
4. A phosphinothricin-resistant plant comprising a plurality of the phosphinothricin-resistant transgenic plant cells of claim 1 , wherein the plant is resistant to glufosinate when applied at a rate of 560 g ai/ha.
5. A seed of the plant of claim 4 .
6. The phosphinothricin-resistant transgenic plant cell of claim 1 , wherein the cell further comprises an insect-resistance gene derived from an organism selected from the group consisting of Bacillus thruingiensis, Plotorhabdus , and Xenorhabdus.
7. The phosphinothricin-resistant transgenic plant cell of claim 1 , wherein the cell further comprises a second herbicide-resistance gene.
8. The phosphinothricin-resistant plant of claim 4 , wherein the plant further comprises a herbicide resistance gene selected from the group consisting of AAD-1 and AAD-12, and wherein the plant is resistant to 2,4-D.
9. The phosphinothricin-resistant plant of claim 4 , wherein the plant further comprises an insect-resistance gene from an organism selected from the group consisting of Bacillus thruingiensis, Photorhabdus , and Xenorhabdus.
10. The phosphinothricin-resistant plant of claim 4 , wherein the plant further comprises a gene for an agronomic trait selected from the group consisting of fungal resistance, stress tolerance, increased yield, improved oil profile, improved fiber quality, viral resistance, delayed ripening, cold tolerance, and salt tolerance.
11. The phosphinothricin-resistant plant of claim 4 , wherein the plant is soybean.
12. The phosphinothricin-resistant plant of claim 4 , wherein the plant is corn.
13. The phosphinothricin-resistant transgenic plant cell of claim 1 , wherein the plant cell comprises a single copy of the polynucleotide of SEQ ID NO:3 integrated in the genome of the plant cell.
14. The phosphinothricin-resistant plant of claim 4 , wherein the plant is resistant to glufosinate when applied at a rate of 1,120 g ai/ha.
15. The phosphinothricin-resistant plant of claim 14 , wherein the plant is soybean.
16. The phosphinothricin-resistant plant of claim 14 , wherein the plant is corn.