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AssigneeE.I. du Pont de Nemours and Company
InventorsSteven Gutteridge, Timothy Caspar, Daniel Cordova, James J. Rauh, Yong Tao, Lihong Wu, Rejane M. Smith
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The genes encoding ryanodine receptor homologs have been characterized from multiple insect families including lepidopteran tobacco budworm (Heliothis virescens), homopteran green peach aphid (Myzus persicae), corn plant hopper (Peregrinus maidis), cotton melon aphid (Aphis gossypii) and fruitfly (Drosophila melanogaster). The full-length genes have been isolated, cloned and amplified in bacterial cells. Expression in insect cells shows that the recombinant protein folds into a functional calcium release channel. The genes and their corresponding polypeptides have a number of uses including, but not limited to, the isolation of other pest ryanodine receptors, the development of screens to identify insecticidally active compounds, use of fragments of genes as pesticides, fragments of protein for antibody production, fragments of protein for determination of the structure of insecticide binding sites, and identification of insecticides that disrupt the calcium balance in cells through other messengers that interact with the receptor calcium release mechanism. Methods are outlined for overcoming toxic effects of expressing recombinant proteins in host cells.
An isolated nucleotide fragment comprising:
An isolated nucleic acid fragment comprises a nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 127, 129, 143, or 145.
A method to isolate nucleic acid fragments encoding ryanodine receptors and related polypeptides, comprising:
An isolated polypeptide having ryanodine receptor activity, wherein the amino acid sequence of the polypeptide and the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 128, 130, 144, or 146, have at least 80% identity.
An isolated nucleic acid fragment encoding an insect ion channel comprising at least two polypeptide sequences set forth in any of SEQ ID NOs:63-119 provided that said polypeptide sequences do not comprise any of SEQ ID NOs:
A method for identifying a nucleic acid sequence encoding an insect ion channel comprising:
A method for expressing an isolated nucleic acid fragment encoding a toxic insect ion channel which comprises:
A method for expressing an isolated nucleic acid fragment encoding a toxic insect ion channel which comprises:
A method for expressing an isolated nucleic acid fragment encoding a toxic insect ion channel which comprises:
A method for expressing an isolated nucleic acid fragment encoding a toxic insect ion channel which comprises:
A recombinant construct comprising in the 5′ to 3′ direction a promoter operably linked to an isolated nucleic acid fragment encoding a toxic insect ion channel
A recombinant construct comprising in the 5′ to 3′ direction a promoter operably linked to an isolated nucleic acid fragment encoding a toxic insect ion channel
A recombinant construct comprising in the 5′ to 3′ direction a promoter operably linked to an isolated nucleic acid fragment encoding a toxic insect ion channel
A recombinant construct comprising in the 5′ to 3′ direction a promoter operably linked to an isolated nucleic acid fragment encoding a toxic insect ion channel wherein said isolated nucleic acid fragment also comprises an intron which interferes with expression of the toxic insect ion channel.
Dependencies are inferred from the claim text.