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Compositions and methods for controlling insects

A composition and technology for insects, applied in the field of compositions and methods for controlling insects, can solve the problems of different insect functions, difficult identification of compounds, vertebrate heart poisoning, etc.

Active Publication Date: 2006-07-26
TYRATECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dimeform and the related formamidines are known to act on octopamine receptors in insects, but have been withdrawn from the market due to their cardiotoxic potential in vertebrates, carcinogenicity in animals and variable action on different insects
Other compounds with less toxicity to mammals and other non-target species are often difficult to identify

Method used

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  • Compositions and methods for controlling insects
  • Compositions and methods for controlling insects
  • Compositions and methods for controlling insects

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] Preparation of Schneider cell line stably transfected with tyramine receptor (TyrR)

[0121] A. PCR amplification and subcloning of Drosophila melanogaster tyramine receptors

[0122] Tyramine receptors were amplified from the head cDNA phage library GH of Drosophila melanogaster obtained from the Berkeley Drosophila Genome Project (Baumann, A., 1999, Drosophila melanogaster mRNA for octopamine receptor, splice variant 1B NCBI direct submission, AccessionAJ007617) . The nucleic acid sequence and peptide sequence of TyrR are as Figure 31A and 31B shown. Phage DNA was purified from liquid culture lysates of this library. (Baxter, et al., 1999, Insect Biochem Mol Biol 29, 461-467). Open reading frame for amplification of the Drosophila tyramine receptor (TyrR) (Han, et al., 1998, J Neurosci 18, 3650-3658; von Nickisch-Rosenegk, et al., 1996. Insect BiochemMol Biol 26, 817-827) is a 5' oligonucleotide: 5' gccgaattc gc caccATGCCATCGGCAGATCAGATCCTG 3' and 3' oligos...

Embodiment 2

[0140] Effects of Treatment of Cells Expressing Tyramine Receptors and Compositions on Intracellular [cAMP]

[0141] Cells were cultured on plates and the medium was changed the day before treatment. When the cells reached approximately 95% confluency, the culture medium was aspirated and replaced with 5ml insect saline (170mM NaCl, 6.0mM KCl, 2.0mM NaHCO 3 , 17.0mM Glucose, 6.0mM NaH 2 PO 4 , 2.0 mM CaCl 2 , 4.0 mM MgCl 2 ; pH 7.0) to wash the cells once. Add approximately 20ml of insect saline, and gently scrape to collect the cells. An aliquot of cells was counted with a hemocytometer, and the cells were centrifuged at 1000 RPM for approximately 5 minutes. Resuspend cells to 3 x 10 6 cells / ml. IBMX was added to a concentration of about 200 µM. Then about 1ml of the cell suspension was aliquoted for processing. Forskolin (cAMP inducing reagent), tyramine or different candidate combinations are added and the cells are incubated at about 27°C for about 10 minutes.

[...

Embodiment 3

[0145] Treatment of Cells Expressing Tyramine Receptors and Compositions for Intracellular [Ca 2+ ]Effect

[0146] The intracellular calcium ion concentration was determined by the fluorescent indicator furan-2 acetoxymethyl (AM) ester ([Ca 2+ ] i) (Enan, et al., Biochem. Pharmacol vol 51, 447-454). In this study, cells expressing tyramine were cultured under standard conditions. Zhou detection buffer (140mM NaCl, 10mM HEPES, 10mM glucose, 5mM KCl, 1mM CaCl 2 , 1 mM MgCl 2 ) to prepare cell suspension, adjust the number of cells to approximately 2×10 per ml 6 cells. Briefly, approximately 1.0ml of the suspension (approximately 2×10 6 cells) were incubated with 5 μM furan 2 / AM for about 30 minutes at about 28°C. After incubation, cells were pelleted by centrifugation at 3700 rpm for approximately 10 seconds at room temperature, and then resuspended in approximately 1.5 ml of assay buffer. [Ca 2+ ]i changes. The excitation wavelength is about 340nm (by binding Ca 2+ p...

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Abstract

The present invention comprises compositions, methods and cell lines related to controlling insects. An embodiment of a composition comprises a plant essential oil and targets at least one receptor of insects chosen from tyramine receptor, Or83b olfactory receptor, and Or43a olfactory receptor, resulting in a change in the intracellular levels of cAMP, Ca2+, or both in the insects.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application Serial No. 60 / 465,320, filed April 24, 2003, and US Provisional Application Serial No. 60 / 532,503, filed December 24, 2003, which are hereby incorporated by reference in their entirety. field of invention [0003] The present invention relates to compositions, methods, cell lines and reporters related to the control of insects. Background of the invention [0004] Animals have chemosensory and mechanosensory systems, which can recognize a variety of environmental stimuli and produce behavioral responses. Behavioral studies have been performed to understand the genetics of these systems. The olfactory system plays an important role in the survival and life of some species, including insects. [0005] Drosophila is a model organism for the study of sensory systems due to its ability to perform mutational analysis by molecular techniques, conduct behavioral ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/00
CPCA01N65/00A01N65/08A01N65/10A01N65/22A01N65/44Y02A50/30A01N27/00A01N31/08A01N31/16A01N35/02A01N37/18A01N49/00A01N65/32A01N53/00A01N65/36
Inventor 伊塞姆·伊那恩
Owner TYRATECH
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