Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
2000-01-11
2002-05-14
Carlson, Karen Cochrane (Department: 1653)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S007100, C530S350000, C530S380000
Reexamination Certificate
active
06387637
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to genes isolated from Arabidopsis that code for proteins essential for seedling growth. The invention also includes the methods of using these proteins as an herbicide target, based on the essentiality of the gene for normal growth and development. The invention is also useful as a screening assay to identify inhibitors that are potential herbicides. The invention may also be applied to the development of herbicide tolerant plants, plant tissues, plant seeds, and plant cells.
BACKGROUND OF THE INVENTION
The use of herbicides to control undesirable vegetation such as weeds in crop fields has become almost a universal practice. The herbicide market exceeds 15 billion dollars annually. Despite this extensive use, weed control remains a significant and costly problem for farmers.
Effective use of herbicides requires sound management. For instance, the time and method of application and stage of weed plant development are critical to getting good weed control with herbicides. Since various weed species are resistant to herbicides, the production of effective new herbicides becomes increasingly important. Novel herbicides can now be discovered using high-throughput screens that implement recombinant DNA technology. Metabolic enzymes found to be essential to plant growth and development can be recombinantly produced through standard molecular biological techniques and utilized as herbicide targets in screens for novel inhibitors of the enzyme activity. The novel inhibitors discovered through such screens may then be used as herbicides to control undesirable vegetation.
Herbicides that exhibit greater potency, broader weed spectrum, and more rapid degradation in soil can also, unfortunately, have greater crop phytotoxicity. One solution applied to this problem has been to develop crops that are resistant or tolerant to herbicides. Crop hybrids or varieties tolerant to the herbicides allow for the use of the herbicides to kill weeds without attendant risk of damage to the crop. Development of tolerance can allow application of a herbicide to a crop where its use was previously precluded or limited (e.g. to pre-emergence use) due to sensitivity of the crop to the herbicide. For example, U.S. Pat. No. 4,761,373 to Anderson et al. is directed to plants resistant to various imidazolinone or sulfonamide herbicides. An altered acetohydroxyacid synthase (AHAS) enzyme confers the resistance. U.S. Pat. No. 4,975,374 to Goodman et al. relates to plant cells and plants containing a gene encoding a mutant glutamine synthetase (GS) resistant to inhibition by herbicides that were known to inhibit GS, e.g. phosphinothricin and methionine sulfoximine. U.S. Pat. No. 5,013,659 to Bedbrook et al. is directed to plants expressing a mutant acetolactate synthase that renders the plants resistant to inhibition by sulfonylurea herbicides. U.S. Pat. No. 5,162,602 to Somers et al. discloses plants tolerant to inhibition by cyclohexanedione and aryloxyphenoxypropanoic acid herbicides. The tolerance is conferred by an altered acetyl coenzyme A carboxylase (ACCase).
Notwithstanding the above described advancements, there remain persistent and ongoing problems with unwanted or detrimental vegetation growth (e.g. weeds). Furthermore, as the population continues to grow, there will be increasing food shortages. Therefore, there exists a long felt, yet unfulfilled need, to find new, effective, and economic herbicides.
SUMMARY OF THE INVENTION
One object of the present invention is to provide an essential gene in plants for assay development for inhibitory compounds with herbicidal activity. Genetic results show that when the 245 gene, the 5283 gene, the 2490 gene, the 3963 gene or the 4036 gene is mutated in Arabidopsis, the resulting phenotype is seedling lethal in the homozygous state. This suggests a critical role for the gene product encoded by the mutated gene.
Using T-DNA insertion mutagenesis, the inventors of the present invention have demonstrated that the activity encoded by the Arabidopsis 245 gene, the Arabidopsis 5283 gene, the Arabidopsis 2490 gene, the Arabidopsis 3963 gene or the Arabidopsis 4036 gene (herein referred to as 245, 5283, 2490, 3963 or 4036 activity) is essential in Arabidopsis seedlings. This implies that chemicals that inhibit the function of the protein in plants are likely to have detrimental effects on plants and are potentially good herbicide candidates. The present invention therefore provides methods of using a purified protein encoded by the gene sequences described below to identify inhibitors thereof, which can then be used as herbicides to suppress the growth of undesirable vegetation, e.g. in fields where crops are grown, particularly agronomically important crops such as maize and other cereal crops such as wheat, oats, rye, sorghum, rice, barley, millet, turf and forage grasses, and the like, as well as cotton, sugar cane, sugar beet, oilseed rape, and soybeans.
The present invention discloses a nucleotide sequence derived from Arabidopsis, designated the 245 gene. The nucleotide sequence of the cDNA clone is set forth in SEQ ID NO:1, and the corresponding amino acid sequence is set forth in SEQ ID NO:2. The nucleotide sequence of the partial genomic DNA sequence is set forth in SEQ ID NO:12. The present invention also includes nucleotide sequences substantially similar to those set forth in SEQ ID NO:1. The present invention also encompasses plant proteins whose amino acid sequence are substantially similar to the amino acid sequences set forth in SEQ ID NO:2. Such proteins can be used in a screening assay to identify inhibitors that are potential herbicides.
The present invention further discloses a nucleotide sequence derived from Arabidopsis, designated the 5283 gene. The nucleotide sequence of the cDNA clone is set forth in SEQ ID NO:3, and the corresponding amino acid sequence is set forth in SEQ ID NO:4. The nucleotide sequence of the genomic DNA sequence is set forth in SEQ ID NO:14. The present invention also includes nucleotide sequences substantially similar to those set forth in SEQ ID NO:3. The present invention also encompasses plant proteins whose amino acid sequence are substantially similar to the amino acid sequences set forth in SEQ ID NO:4. Such proteins can be used in a screening assay to identify inhibitors that are potential herbicides.
The present invention further discloses a nucleotide sequence derived from Arabidopsis, designated the 2490 gene. The nucleotide sequence of the cDNA clone is set forth in SEQ ID NO:5, and the corresponding amino acid sequence is set forth in SEQ ID NO:6. The nucleotide sequence of the genomic DNA sequence is set forth in SEQ ID NO:19. The present invention also includes nucleotide sequences substantially similar to those set forth in SEQ ID NO:5. The present invention also encompasses plant proteins whose amino acid sequence are substantially similar to the amino acid sequences set forth in SEQ ID NO:6. Such proteins can be used in a screening assay to identify inhibitors that are potential herbicides.
The present invention further discloses a nucleotide sequence derived from Arabidopsis, designated the 3963 gene. The nucleotide sequence of the cDNA clone is set forth in SEQ ID NO:7, and the corresponding amino acid sequence is set forth in SEQ ID NO:8. The nucleotide sequence of the genomic DNA sequence is set forth in SEQ ID NO:24, which contains genomic DNA sequences from both the portion of the MDK4 clone annotated as MDK4.6 and added sequences on the 3′ end based on the inventors' reported cDNA clone. The present invention also includes nucleotide sequences substantially similar to those set forth in in SEQ ID NO:7. The present invention also encompasses plant proteins whose amino acid sequence are substantially similar to the amino acid sequences set forth in SEQ ID NO:8. Such proteins can be used in a screening assay to identify inhibitors that are potential herbicides.
The present invention further discloses a nucleotide sequence derived from Arabidopsis
Budziszewski Gregory J.
Levin Joshua Z.
Potter Sharon L.
Wegrich Lynette M.
Carlson Karen Cochrane
Lebel Edouard G.
Meigs J. Timothy
Robinson Hope A.
Stults Larry W.
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