1. A method for detecting the activity of GPR86 in a sample comprising the steps of:
a) incubating a sample comprising GPR86 and ADP under conditions which permit binding of GRP86 and ADP, and
b) detecting a second messenger.
2. The method of claim 1 further comprising the steps of:
a) incubating a second sample comprising GPR86 in the absence of ADP under conditions which permit binding of GRP86 and ADP, and
b) detecting a second messenger.
3. The method of claim 1 wherein said sample comprises cells expressing GPR86.
4. The method of claim 3 wherein said cells are selected from the group consisting of: COS7-cells, a CHO cell, a LM (TK-) cell, a NIH-3T3 cell, HEK-293 cell, K-562 cell, a 1321N1 astrocytoma cell and other cell lines.
5. The method of claim 1 wherein said sample comprises cell membranes bearing GPR86.
6. The method of claim 1, wherein step a) is further performed in the presence of G16 polypeptide.
7. A method of screening for a candidate modulator of GPR86 activity using cells expressing GPR86, said method compromising:
a) incubating a first sample of said cells in the presence of said candidate modulator and a second sample of said cells in the absence of said candidate modulator, both said samples under conditions which permit binding of ADP to GPR86;
b) detecting a signalling activity of GPR86 polypeptide in said first and second samples, and
c) comparing the results of said second messenger assays for said first and second samples.
8. The method of claim 7 wherein said cell is selected from the group consisting of: COS7-cells, a CHO cell, a LM (TK-) cell, a NIH-3T3 cell, HEK-293 cell, K-562 cell, a 1321N1 astrocytoma cell and other cell lines.
9. A method of screening for a candidate modulator of GPR86 activity using cell membranes bearing GPR86, said method comprising:
a) incubating a first sample of said cell membranes in the presence of said candidate modulator and a second sample of said cell membranes in the absence of said candidate modulator, both said samples under conditions which permit binding of ADP to GPR86;
b) detecting a signalling activity of GPR86 polypeptide in said first and second samples, and
c) comparing the results of said second messenger assays for said first and second samples.
10. The methods of claim 7 or 9, further performed in the presence of G 16 polypeptide.
11. A method for determining if a candidate modulator increases or decreases the activity of GPR86 using cells expressing GPR86, said method comprising:
a) incubating a first sample of said cells in the presence of said candidate modulator and a second sample of said cells in the absence of said candidate modulator, both said samples under conditions which permit binding of ADP to GPR86;
b) detecting a signalling activity of GPR86 polypeptide in said first and second samples, and
c) comparing the results of said second messenger assays for said first and second samples.
12. The method of claim 11 wherein said cells are selected from the group consisting of: COS7-cells, a CHO cell, a LM (TK-) cell, a NIH-3T3 cell, HEK-293 cell, K-562 cell and a 1321N1 astrocytoma cell and other cell lines.
13. A method for determining if a candidate modulator increases or decreases the activity of GPR86 using cell membranes bearing GPR86, said method comprising:
a) incubating a first sample of said cell membranes in the presence of said candidate modulator and a second sample of said cell membranes in the absence of said candidate modulator, both said samples under conditions which permit binding of ADP to GPR86;
b) detecting a signalling activity of GPR86 polypeptide in said first and second samples, and
c) comparing the results of said second messenger assays for said first and second samples.
14. The methods of claim 11 or 13, further performed in the presence of G16 polypeptide.
15. A kit for detecting binding to GPR86 or a modulator of GPR86, said kit comprising GPR86 and ADP, and packaging materials therefore, wherein said GPR86 and ADP are packaged separately.
16. The kit of claim 15, wherein GPR86 is present in a cell.
17. The kit of claim 16 wherein said cell is selected from the group consisting of: COS7-cells, a CHO cell, a LM (TK-) cell, a NIH-3T3 cell, HEK-293 cell, K-562 cell and a 1321N1 astrocytoma cell and other cell lines.
18. The kit of claim 15, wherein GPR86 is associated with a cell membrane.
19. The kit of claim 15, said kit further comprising the components of a second messenger assay.
20. The kit of claim 15, wherein the modulator is detected using an antibody specific for GPR86 or a GPR86-specific nucleic acid probe.
21. The kit of claim 15, said kit further comprising G16 polypeptide.
22. A non-human mammal comprising a partial or total deletion of the ortholog sequence of the human polynucleotide (SEQ ID NO.1), preferably a non-human mammal comprising a homologous recombinant knockout of said polynucleotide or a transgenic non-human mammal overexpressing above natural level said polynucleotide.
23. A method of identifying an agent that modulates the function of GPR86, said method comprising:
a) contacting a GPR86 polypeptide in the presence and absence of a candidate modulator under conditions permitting the binding of said ADP to said GPR86 polypeptide; and
b) measuring the binding of said GPR86 polypeptide to said candidate modulator, relative to the binding in the absence of said candidate modulator, identifies said candidate modulator as an agent that modulates the function of GPR86.
24. The method of claim 23 wherein said measuring is performed using a method selected from label displacement, surface plasmon resonance, fluorescence resonance energy transfer, fluorescence quenching, and fluorescence polarization.
25. The method of any one of claims 7, 9, 11 and 13 wherein said candidate modulator is selected from the group consisting of a natural or synthetic peptide, a polypeptide, an antibody or antigen-binding fragment thereof, a lipid, a carbohydrate, a nucleic acid, and a small organic molecule.
26. The method of any one of claims 7, 9, 11 and 13 wherein said step of measuring a signalling activity of said GPR86 polypeptide comprises detecting a change in the level of a second messenger.
27. The method of either of claims 7,9,11 or 13 wherein the step of detecting a signalling activity comprises measurement of guanine nucleotide binding or exchange, adenylate cyclase activity, cAMP, protein kinase C activity, phosphatidylinosotol breakdown, diacylglycerol, inositol triphosphate, intracellular calcium, arachinoid acid concentration, MAP kinase activity, tyrosine kinase activity, reporter gene expression.
28. The method of claims 27 wherein said measuring a signalling activity comprises using an aequorin-based assay.
29. A method of modulating the activity of a GPR86 polypeptide in a cell, said method comprising the step of delivering to said cell an agent that modulates the activity of a GPR86 polypeptide, such that the activity of GPR86 is modulated.
30. A method of diagnosing a disease or disorder characterized by detecting dysregulation of GPR86 signalling, said method comprising:
a) contacting a tissue sample with an antibody specific for a GPR86 polypeptide;
b) detecting binding of said antibody to said tissue sample; and
c) comparing the binding detected in step (b) with a standard, wherein a difference in binding relative to said standard is diagnostic of a disease or disorder characterized by dysregulation of GPR86.
31. A method of diagnosing a disease or disorder characterized by dysregulation of GPR86 signalling, said method comprising:
a) contacting a tissue sample with an antibody specific for a GPR86 specific ligand;
b) detecting binding of said antibody to said tissue sample; and
c) comparing the binding detected in step (b) with a standard, wherein a difference in binding relative to said standard is diagnostic of a disease or disorder characterized by dysregulation of GPR86.
32. A method of diagnosing a disease or disorder characterized by dysregulation of GPR86 signalling, said method comprising:
a) contacting a tissue sample with an antibody specific for a GPR86 polypeptide and an antibody specific for a GPR86-specific ligand;
b) detecting binding of said antibodies to said tissue sample; and
c) comparing the binding detected in step (b) with a standard, wherein a difference in the binding of either antibody or both, relative to said standard, is diagnostic of a disease or disorder characterized by dysregulation of GPR86.
33. A method of diagnosing a disease or disorder characterized by dysregulation of GPR86 signalling, said method comprising:
a) isolating nucleic acid from a tissue sample;
b) amplifying a GPR86 polynucleotide, using said nucleic acid as a template; and
c) comparing the amount of amplified GPR86 polynucleotide produced in step (b) with a standard, wherein a difference in said amount of amplified GPR86 polynucleotide relative to said standard is diagnostic of a disease or disorder characterized by dysregulation of GPR86.
34. A method of diagnosing a disease or disorder characterized by dysregulation of GPPR86 signalling, said method comprising:
a) isolating nucleic acid from a tissue sample;
b) amplifying a GPR86-specific ligand polynucleotide, using said nucleic acid as a template; and
c) comparing the sequence of said amplified GPR86-specific ligand polynucleotide produced in step (b) with a standard, wherein a difference in said sequence, relative to said standard is diagnostic of a disease or disorder characterized by dysregulation of GPR86.
35. The method of claim 33 wherein said GPR86 polynucleotide is SEQ ID NO: 1.
36. The method of claim 33 or 34 wherein said comparing the amount is performed on a microarray.
37. An antibody specific for a GPR86 polypeptide.
38. The method of any one of claims 1-6 wherein said incubating is performed in or on virus-induced budding membranes containing a GPR86 polypeptide. A method of detecting the presence, in a sample, of an agent that modulates the function of GPR86, said method comprising:
a) contacting a GPR86 polypeptide with said sample;
b) measuring a signalling activity of said GPR86 polypeptide in the presence of said sample; and
c) comparing said activity measured in the presence of said sample to said activity measured in a reaction in which said GPR86 polypeptide is contacted with a ADP present at its EC50, wherein an agent that modulates the function of GPR86 is detected if the amount or said activity measured in the presence of said sample is at least 50% of the amount induced by said ADP present at it EC50.
39. A kit for screening for agents that modulate the signalling activity of GPR86, said kit comprising an isolated polynucleotide encoding a GPR86 polypeptide and means for detecting GPR86 signalling, and packaging materials therefore.
40. The kit of claim 39, further comprising the ligand, ADP.
41. A kit for screening for agents that modulate the signalling activity of GPR86, said kit comprising a cell transformed with a polynucleotide encoding a GPR86 polypeptide and means for detecting GPR86 signalling, and packaging materials therefore.
42. The kit of claim 39 or 41, wherein the said agents are detected using an antibody specific for GPR86 or a GPR86-specific nucleic acid probe.
43. A kit for the diagnosis of a disease or disorder characterized by dysregulation of GPR86 signalling, said kit comprising an isolated GPR86 polypeptide and means for detecting GPR86 signalling, and packaging materials therefore.
44. The kit of claim 43, wherein the said diagnosis or disorder is detected using an antibody specific for GPR86 or a GPR86-specific nucleic acid probe.
45. The kit of claims 15, 39, 41 or 43, further comprising a standard of GPR86 activity as measured in a cell line expressing GPR86 in the presence of ADP.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
Claims
1. An improved agricultural seed planter, said planter having a main frame operably supporting a plurality of adjacently spaced row units, each said row units having a row unit frame operably supporting a seed hopper, a furrow opening assembly comprising at least one furrow opening disk adapted to open a seed furrow in a soil surface, a furrow closing assembly adapted to close the seed furrow in the soil surface, and a seed meter disposed a distance above the soil surface and adapted to discharge individual seeds at regular intervals, the improvement comprising:
a seed delivery apparatus adapted to receive the individual seeds discharged from the seed meter and to controllably convey the individually discharged seeds in sequence to the open furrow, said seed delivery apparatus comprising:
a housing in communication with the seed meter to sequentially receive the individually discharged seeds therefrom, said housing adapted to be operably supported by the row unit frame and having an open end disposed in substantial alignment with the open furrow, and further wherein at least a portion of said housing is disposed transversely adjacent said at least one furrow opening disk of the furrow opening assembly;
a seed conveyor disposed in said housing and adapted to individually convey the individual discharged seeds in sequence through said housing at a controlled conveyor speed before being serially released through said open end in said housing and deposited into the open furrow.
2. The improved agricultural seed planter of claim 1 wherein said seed conveyor includes a continuous belt disposed around a drive pulley and an idler pulley, said continuous belt having a predetermined plurality of flights that individually receive the individual discharged seeds from the seed meter and retain the individual seeds in sequence until being serially released through said open end in said housing and deposited into the open furrow.
3. The improved agricultural seed planter of claim 2 wherein said drive pulley is operably coupled to the seed meter and is operably rotatably driven thereby in synchronous relation.
4. The improved agricultural seed planter of claim 1, wherein said seed conveyor includes a first continuous belt having a length disposed in spaced opposition to a length of a second continuous belt, said first continuous belt disposed around a first drive pulley and a first idler pulley, said second continuous belt disposed around a second drive pulley and a second idler pulley, said first drive pulley operably coupled to said second drive pulley such that said opposing lengths of said first and second continuous belts travel at substantially the same speed, said first and second continuous belts disposed within said housing to receive the individual seeds discharged by the seed meter and retain the individual seeds in sequence between their opposing lengths before being serially released through said open end in said housing and deposited into the open furrow.
5. The improved agricultural seed planter of claim 4 wherein at least one of said first and second drive pulleys is operably coupled to the seed meter and is operably rotatably driven thereby.
6. The improved agricultural seed planter of claim 4 wherein at least one of said first and second drive pulleys is operably coupled to a drive mechanism and is operably rotatably driven thereby independent the seed meter.
7. The improved agricultural seed planter of claim 4 wherein said length of at least one of said first and second continuous belts is movably adjustable relative to said opposing length of said other continuous belt to accommodate various sizes of the individual discharged seeds.
8. In combination, a seed meter and a seed delivery apparatus for an agricultural seed planter having a furrow opening assembly comprising at least one furrow opening disk adapted to open a seed furrow in a soil surface, the seed meter adapted to discharge individual seeds at regular intervals, said seed delivery apparatus comprising:
a housing in communication with the seed meter and adapted to serially receive the individually discharged seeds therefrom, and having an open end disposed in substantial alignment with the open furrow in the soil surface, and further wherein at least a portion of said housing is disposed transversely adjacent said at least one furrow opening disk of the furrow opening assembly;
a seed conveyor disposed in said housing and adapted to individually convey the individual discharged seeds in sequence through said housing at a controlled conveyor speed until being serially released through said open end in said housing and dispensed into the open furrow.
9. The combination of claim 8 wherein said seed conveyor includes a continuous belt disposed around a drive pulley and an idler pulley, said continuous belt having a predetermined plurality of flights that individually receive the individual discharged seeds from the seed meter and individually retain the individual seeds until being released through said open end in said housing and dispensed into said open furrow.
10. The combination of claim 9 wherein said drive pulley is operably coupled to the seed meter and is operably rotatably driven thereby in synchronous relation.
11. The combination of claim 8, wherein said seed conveyor includes a first continuous belt having a length disposed in spaced opposition to a length of a second continuous belt, said first continuous belt disposed around a first drive pulley and a first idler pulley, said second continuous belt disposed around a second drive pulley and a second idler pulley, said first drive pulley operably coupled to said second drive pulley such that said opposing lengths of said first and second continuous belts travel at substantially the same speed, said first and second continuous belts disposed within said housing to receive the individual seeds discharged by the seed meter and retain the individual seeds in sequence between their opposing lengths before being serially released through said open end in said housing and deposited into the open furrow.
12. The combination of claim 11 wherein at least one of said first and second drive pulleys is operably coupled to the seed meter and is operably rotatably driven thereby.
13. The combination of claim 11 wherein at least one of said first and second drive pulleys is operably coupled to a drive mechanism and is operably rotatably driven thereby independent the seed meter.
14. The combination of claim 11 wherein said length of at least one of said first and second continuous belts is movably adjustable relative to said opposing length of said other continuous belt to accommodate various sizes of the individual discharged seeds.
15. A method of improving accuracy in placement of individual seeds in an open seed furrow in a soil surface, said method comprising:
providing a furrow opening assembly having at least one furrow opening disk;
providing a seed meter disposed a distance above the soil surface and adapted to discharge individual seeds at regular intervals;
providing a housing in communication with said seed meter adapted to receive the individual discharged seeds in sequence therefrom, said housing having an open end proximate the soil surface in substantial alignment with the open seed furrow, and further wherein at least a portion of said housing is disposed transversely adjacent said at least one furrow opening disk of said furrow opening assembly;
conveying the individual discharged seeds through said housing in sequence at a controlled conveyor speed;
releasing the individual conveyed seeds in sequence through said open end in said housing near the soil surface and into the open seed furrow.
16. The method of claim 15 wherein said step of conveying the individual discharged seeds through said housing includes providing a continuous belt disposed around a drive pulley and an idler pulley, said continuous belt having a predetermined plurality of flights that individually receive the individual seeds and individually retain the individual seeds in sequence through said housing before being serially released through said open end in said housing and deposited into the open seed furrow.
17. The method of claim 16 wherein said drive pulley is operably coupled to the seed meter and is operably rotatably driven thereby in synchronous relation.
18. The method of claim 15 wherein said step of conveying the individual discharged seeds through said housing includes providing a first continuous belt having a length disposed in spaced opposition to a length of a second continuous belt, said first continuous belt disposed around a first drive pulley and a first idler pulley, said second continuous belt disposed around a second drive pulley and a second idler pulley, said first drive pulley operably coupled to said second drive pulley such that said opposing lengths of said first and second continuous belts travel at substantially the same speed, said first and second continuous belts disposed within said housing to receive the individual seeds discharged by the seed meter and retain the individual seeds in sequence between their opposing lengths before being serially released through said open end in said housing and deposited into the open seed furrow.
19. The method of claim 18 wherein at least one of said first and second drive pulleys is operably coupled to the seed meter and is operably rotatably driven thereby.
20. The method of claim 18 wherein at least one of said first and second drive pulleys is operably coupled to a drive mechanism and is operably rotatably driven thereby independent the seed meter.
21. method of claim 18 wherein said length of at least one of said first and second continuous belts is movably adjustable relat ive to said opposing length of said other continuous belt to accommodate various sizes of the individually discharged seeds.
22. A seed delivery device for an agricultural planter, comprising:
a conveyor support structure having a first end positioned near a seed metering device to receive seeds metered individually and in a sequence therefrom, and having a second end positioned proximate an open seed furrow and in substantial alignment therewith, and further wherein at least a portion of said conveyor support structure is disposed transversely adjacent a furrow opening disk; and
a seed conveyor mounted moveably relative to said conveyor support structure and operable at a controlled conveyor speed to transport the seeds in said sequence and at said controlled conveyor speed from said first end toward said second end, said seed conveyor further being operable to serially release the seeds proximate said second end to discharge the seeds in said sequence into said open seed furrow as the agricultural planter traverses the field.
23. The device of claim 22 wherein said controlled conveyor speed is constant.
24. The device of claim 22 wherein said controlled conveyor speed is variable in proportion to a linear speed of the agricultural planter carrying said conveyor support structure.
25. The device of claim 22 wherein said controlled conveyor speed is variable in proportion to a metering speed of said metering device.
26. The device of claim 22 wherein said conveyor support structure comprises a housing substantially enclosing said seed conveyor, said housing including a seed receiving opening near said first end, and a seed dispensing opening near said second end.
27. The device of claim 26 wherein said seed conveyor comprises an endless belt and a plurality of flights extended outwardly from said belt and substantially uniformly spaced apart from one another.
28. The device of claim 22 further including a seed guide near said second end of said conveyor support structure, disposed to encounter the seeds dispensed proximate said second end.
29. The device of claim 22 wherein said seed conveyor comprises first and second endless belts including respective first and second belt segments extending between said first and second ends, confronting one another, and spaced apart from one another by a distance less than a diameter of the seeds whereby said endless belts, when moved in concert at said controlled conveyor speed, are adapted to transport the seeds toward said second end at said controlled conveyor speed.
30. A seed transporting apparatus for an agricultural planter, comprising:
a conveyor support structure adapted for an attachment to a seed metering device of an agricultural planter, with a first end of said conveyor support structure positioned near said metering device to receive seeds metered individually and in a sequence therefrom, and with a second end of said conveyor support structure relatively remote with respect to said metering device;
a first flexible endless belt mounted moveably relative to said conveyor support structure; and
a second flexible endless belt mounted moveably relative to said conveyor support structure;
wherein said first and second endless belts include respective first and second belt segments disposed in substantially parallel and confronting relation, moveable in concert at a controlled conveyor speed from said first end toward said second end;
wherein said first and second belts along the first and second belt segments are spaced apart from one another by a distance less than a diameter of the seeds, and thereby are adapted to releasably hold the seeds fixed relative to said belts for travel toward said second end at said controlled conveyor speed and in said sequence; and
wherein said first and second belts diverge from one another near said second end, to release the seeds in said sequence.
31. The apparatus of claim 30 further including a belt tensioning component for adjusting tension throughout at least one of said first and second endless belts.
32. The apparatus of claim 31 wherein said belt tensioning component includes a first tensioning member operable to adjust said tension throughout said first endless belt and a second tensioning member operable to adjust said tension throughout said second endless belt.
33. The apparatus of claim 30 wherein at least one of said first and second endless belts is mounted adjustably relative to said conveyor support structure to facilitate changing said selected distance to accommodate different sizes of the seeds.
34. The apparatus of claim 30 further including a first drive pulley mounted rotatably with respect to said conveyor support structure and engaged with said first endless belt, and a second drive pulley mounted rotatably relative to said conveyor support structure and drivably engaged with said second endless belt.
35. The apparatus of claim 34 wherein said first drive pulley and said second drive pulley are drivably associated with one another for operation in concert.
36. The apparatus of claim 30 wherein said second end of said conveyor support structure, when said first end of the conveyor support structure is positioned near the metering device, is disposed proximate an open furrow in an agricultural field for movement along the furrow as an agricultural planter carrying said conveyor support structure traverses said field.
37. The apparatus of claim 30 further including a seed metering device coupled to said conveyor support structure for supplying seeds metered individually and in a sequence to said endless belts near said first end of said conveyor support structure.
38. The apparatus of claim 30 further including a furrow opening assembly, a gauge assembly for vertically positioning said furrow opening assembly, a furrow closing assembly, a seed hopper, and said seed metering device mounted with respect to said conveyor support structure and cooperating with said conveyor support structure to provide a row unit in which said furrow opening assembly is disposed forwardly of the endless belts and at least partially below a ground surface, said furrow closing assembly is disposed behind said endless belts, and said second end of said conveyor support structure is disposed proximate said ground surface.