Methods of screening compositions for potential insect control activity
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, Or83 b olfactory receptor, and Or43 a olfactory receptor, resulting in a change in the intracellular levels of cAMP, Ca2+, or both in the insects.
1. A method of screening a composition for potential insect control activity, comprising:
providing insect cells expressing olfactory receptor Or83b and a tyramine receptor;
adding said composition to the cells;
measuring the binding affinity of said composition to the receptor; and
selecting a composition having an affinity for either of said receptors, wherein said affinity is indicative of potential insect control activity.
2. A method of screening a composition for potential insect control activity comprising:
providing insect cells expressing olfactory receptor Or83b and a tyramine receptor;
adding said composition to the cells;
extracting intracellular cAMP or Ca 2+ from the cells;
measuring the intracellular cAMP or Ca 2+ levels;
comparing the intracellular cAMP or Ca 2+ levels in cells treated with said composition to the intracellular cAMP or Ca 2+ levels in untreated cells; and
selecting a composition that causes a change in intracellular cAMP, Ca 2+ , or both via at least one of said receptors, wherein said change is indicative of potential insect control activity.
3. The method of claim 2 , wherein the tyramine receptor is expressed from the nucleic acid sequence as set for the in SEQ ID NO:1.
4. The method of claim 2 , wherein the olfactory receptor Or83b is expressed from the nucleic acid sequence as set forth in SEQ ID NO:5.
5. The method of claim 2 , wherein intracellular cAMP is extracted from the cells and its levels are measured and compared.
6. The method of claim 2 , wherein intracellular Ca 2+ is extracted from the cells and its levels are measured and compared.
7. A method of screening a composition for potential insect control activity, comprising:
providing insect cells expressing olfactory receptor 83b and a tyramine receptor;
adding said composition to the cells;
measuring a characteristic of the cells in the presence of the composition, wherein the characteristic results from an interaction between the composition and at least one of the said receptors and is indicative of a potential for insect control activity; and
selecting a composition suitable for insect control based on the results of the measuring step.
8. The method of claim 7 , wherein the characteristic is selected from the group consisting of: a binding affinity of the composition to the receptor; a change in intracellular cAMP level; and a change in intracellular Ca 2+ level.
9. The method of claim 1 , wherein the insect cells additionally express olfactory receptor 43a.
10. The method of claim 2 , wherein the insect cells additionally express olfactory receptor 43a.
11. The method of claim 7 , wherein the insect cells additionally express olfactory receptor 43a.