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  Nucleic acid molecules encoding novel bacillus thuringiensis cry2ae insecticidal proteins, plant cells, plant or seeds comprising the nucleic acid molecules and methods of using sameUSPTO Application #: 20080047034Title: Nucleic acid molecules encoding novel bacillus thuringiensis cry2ae insecticidal proteins, plant cells, plant or seeds comprising the nucleic acid molecules and methods of using same Abstract: The invention pertains to novel insecticidal compounds derived from Bacillus thuringiensis strains. New proteins designated Cry2Ae, Cry2Af, and Cry2Ag, and variants thereof are provided, as well as DNA sequences encoding these proteins or their variants. Further provided are recombinant hosts expressing such proteins, particularly plant cells and plants. (end of abstract) Agent: Hunton & Williams LLP Intellectual Property Department - Washington, DC, US Inventors: Greta Arnaut, Annemie Boets, Stijn Vanneste, Jeroen Van Rie, Sara Van Houdt USPTO Applicaton #: 20080047034 - Class: 800279000 (USPTO) Related Patent Categories: Multicellular Living Organisms And Unmodified Parts Thereof And Related Processes, Method Of Introducing A Polynucleotide Molecule Into Or Rearrangement Of Genetic Material Within A Plant Or Plant Part, The Polynucleotide Confers Pathogen Or Pest Resistance The Patent Description & Claims data below is from USPTO Patent Application 20080047034. Brief Patent Description - Full Patent Description - Patent Application Claims
[0001] The present application claims priority to application Ser. No. 09/756,296, the disclosure of which is hereby incorporation by reference. BACKGROUND OF THE INVENTION [0002] (i) Field of the Invention [0003] The present invention relates to new nucleic acid sequences, particularly DNA sequences, encoding insecticidal proteins produced by Bacillus thuringiensis strains. Particularly, new nucleic acid sequences, particularly DNA sequences encoding proteins designated as Cry2Ae, Cry2Af and Cry2Ag are provided which are useful to protect plants from insect damage. Also included herein are micro-organisms and plants transformed with a nucleic acid sequence, particularly a DNA sequence, encoding at least one of the newly isolated Cry2A proteins. [0004] (ii) Description of Related Art [0005] Bacillus thuringiensis (abbreviated herein as "Bt") is well known for its specific toxicity to insect pests, and has been used since almost a century to control insect pests of plants. In more recent years, transgenic plants expressing Bt proteins were made which were found to successfully control insect damage on plants (e.g., Vaeck et al., 1987, Jansens et al., 1997). [0006] Despite the isolation of quite a number of insecticidal Bt genes, the search for new genes encoding insecticidal proteins continues. Indeed, insecticidal Bt proteins are known to have a relatively narrow target insect range compared to chemical insecticides. Also, having multiple toxins to the same target insect species allows the use of proteins having different modes of action so that insect resistance development can be prevented or delayed. And, insecticidal Bt proteins with different amino acid sequences have different levels of insecticidal efficacy against specific insects, making it desirable to have several different insecticidal proteins available in order to be able to control the relevant insect pests of different crop plants. [0007] Previously, several types of Cry2A-proteins were identified (see Crickmore et al., 1998, incorporated herein by reference). [0008] The new Cry2Ae protein of this invention has the highest amino acid sequence identity to the Cry2Aa1 protein (Donovan et al., GenBank accession number M31738), but still differs in about 9 percent of its amino acid sequence. [0009] The closest sequence identity to the Cry2Af protein was found in the Cry2Ab1 protein (Widner and Whiteley, GenBank accession number M23724), but both proteins still differ in about 5 percent of their amino acid sequence. [0010] The closest sequence identity to the Cry2Ag protein was found in the Cry2Ac1 protein (Wu et al., GenBank accession number X57252), but both proteins still differ in about 20 percent of their amino acid sequence. [0011] Further known Cry2A proteins include the Cry2Ad1 protein (Choi et al., 1999), and other Cry2Aa, Cry2Ab, and Cry2Ac proteins (Crickmore et al., 1998). Cry2A-like proteins and DNA sequences encoding them are also shown in U.S. Pat. No. 5,338,544, in published PCT patent application WO 00/26371 and in published PCT patent application WO 98/40490. [0012] Expression of Cry2A-type proteins in plants has been described, e.g., in Kota et al. (1999) and in published PCT patent application WO 00/26371. SUMMARY AND OBJECTS OF THE INVENTION [0013] In accordance with this invention, there is provided a nucleic acid sequence, particularly a DNA sequence, encoding a protein comprising the amino acid sequence selected from the group consisting of: a) the amino acid sequence of the smallest toxic fragment of the protein encoded by the cry2Ae gene deposited at the BCCM-LMBP under accession number LMBP 4248, b) the amino acid sequence of the smallest toxic fragment of the protein encoded by the cry2Af gene deposited at the BCCM-LMBP under accession number LMBP 4247, and c) the amino acid sequence of the smallest toxic fragment of the protein encoded by the cry2Ag gene deposited at the BCCM-LMBP under accession number LMBP 4249. [0014] Particularly preferred in accordance with this invention is a nucleic acid sequence, particularly a DNA sequence, encoding a protein comprising the amino acid sequence selected from the group consisting of: the amino acid sequence of an insecticidal fragment of the protein of SEQ ID No. 2, the amino acid sequence of an insecticidal fragment of the protein of SEQ ID No. 4, the amino acid sequence of an insecticidal fragment of the protein of SEQ ID No. 4. [0015] Further, in accordance with this invention are provided nucleic acid sequences, particularly DNA sequences, encoding a protein comprising the amino acid sequence selected from the group consisting of: the amino acid sequence of SEQ ID No. 2 from amino acid position 1 to amino acid position 632, the amino acid sequence of SEQ ID No. 4 from amino acid position 1 to amino acid position 632, and the amino acid sequence of SEQ ID No. 6 from amino acid position 1 to amino acid position 627. [0016] Further, in accordance with this invention are provided the above nucleic acid sequences, particularly DNA sequences, comprising an artificial sequence, having a different codon usage compared to the naturally occurring sequence, but encoding the same protein or its insecticidal fragment, preferably such codon usage resembles that of plants, particularly the host plant in which the nucleic acid sequence, particularly the DNA, is to be transformed. [0017] Even further provided in accordance with this invention is a protein comprising the amino acid sequence selected from the group consisting of: a) the amino acid sequence of the smallest toxic fragment of the protein encoded by the cry2Ae gene deposited at the BCCM-LMBP under accession number LMBP 4248, b) the amino acid sequence of the smallest toxic fragment of the protein encoded by the cry2Af gene deposited at the BCCM-LMBP under accession number LMBP 4247, and c) the amino acid sequence of the insecticidal smallest toxic fragment of the protein encoded by the cry2Ag gene deposited at the BCCM-LMBP under accession number LMBP 4249. [0018] Particularly preferred herein is a protein comprising the amino acid sequence selected from the group consisting of: the amino acid sequence of an insecticidal fragment of the protein of SEQ ID No. 2, the amino acid sequence of an insecticidal fragment of the protein of SEQ ID No. 4 , and the amino acid sequence of an insecticidal fragment of the protein of SEQ ID No. 6. [0019] Also provided herein are chimeric genes comprising the DNA as defined above under the control of a plant-expressible promoter, and plant cells, plants or seeds transformed to contain those chimeric genes, particularly plant cells, plants, or seeds selected from the group consisting of: corn, cotton, rice, tobacco, oilseed rape, Brassica species, eggplant, soybean, potato, sunflower, tomato, sugarcane, tea, beans, tobacco, strawberry, clover, cucumber, watermelon, pepper, oat, barley, wheat, dahlia, gladiolus, chrysanthemum, sugarbeet, sorghum, alfalfa, apple, pear, strawberry, and peanut. In accordance with this invention, the chimeric gene can be integrated in the nuclear, plastid or mitochondrial DNA of the plant cells, or can also contain a DNA encoding an effective targeting or transit peptide for targeting to the vacuole, chloroplast, mitochondrium, plastid, or for secretion. [0020] Further in accordance with this invention are provided micro-organisms, transformed to contain any of the above DNA sequences, particularly those. selected from the genus Pseudomonas, Agrobacterium, Escherichia, or Bacillus. [0021] Also provided herein is a process for controlling insects, comprising expressing any of the above nucleic acid sequences, particularly DNA sequences, in a host cell, particularly plant cells, and contacting insects with said host cells, and a process for rendering a plant resistant to insects, comprising transforming plants cells with any of the above DNA sequences or chimeric genes, and regenerating transformed plants from such cells which are resistant to insects. [0022] This invention also relates to a method for controlling lepidopteran insects, particularly lepidopteran insect pests of cotton, corn or soybean, which method comprises applying to an area or plant to be protected, a Cry2A protein as defined herein, preferably a Cry2Ae protein as defined herein, (i.e., by planting a plant transformed with a cry2A gene of this invention, or by spraying a composition containing a Cry2A protein of this invention). The invention also relates to the use of the Cry2A proteins of this invention, particularly the Cry2Ae protein, against Lepidopteran insect pests to minimize damage to soybean plants. Continue reading... Full patent description for Nucleic acid molecules encoding novel bacillus thuringiensis cry2ae insecticidal proteins, plant cells, plant or seeds comprising the nucleic acid molecules and methods of using same Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Nucleic acid molecules encoding novel bacillus thuringiensis cry2ae insecticidal proteins, plant cells, plant or seeds comprising the nucleic acid molecules and methods of using same patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. 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