What is claimed is:
1. An isolated polynucleotide comprising a polynucleotide which is at least 70% identical to a member selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide, or a biologically or immunologically active fragment thereof, comprising the amino acid sequence set forth in FIG. 2 (SEQ ID NO: 2);
(b) a polynucleotide encoding a polypeptide comprising amino acid 21 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2);
(c) a polynucleotide encoding a polypeptide comprising amino acid 27 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2); and
(d) a polynucleotide which is complementary to the polynucleotide of (a), (b), or (c).
2. The polynucleotide of claim 1 wherein the polynucleotide is DNA.
3. The polynucleotide of claim 1 wherein the polynucleotide is RNA.
4. The polynucleotide of claim 1 wherein the polynucleotide is genomic DNA.
5. The polynucleotide of claim 2 wherein the polynucleotide encodes the polypeptide comprising amino acids 1 to 331 as set forth in FIG. 2 (SEQ ID NO: 2).
6. The polynucleotide of claim 2 wherein the polynucleotide encodes the polypeptide comprising amino acid 21 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2).
7. The polynucleotide of claim 2 wherein the polynucleotide encodes the polypeptide comprising amino acid 27 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2).
8. The polynucleotide of claim 1 wherein the polynucleotide comprises the sequence as set forth in FIG. 1 (SEQ ID NO: 1) from nucleotide 1 to nucleotide 1770.
9. The polynucleotide of claim 1 wherein the polynucleotide comprises the sequence as set forth in FIG. 1 (SEQ ID NO: 1) from nucleotide 296 to nucleotide 1291.
10. The polynucleotide of claim 1 wherein the polynucleotide comprises the sequence as set forth in FIG. 1 (SEQ ID NO: 1) from nucleotide 356 to nucleotide 1291.
11. The polynucleotide of claim 1 wherein the polynucleotide comprises the sequence as set forth in FIG. 1 (SEQ ID NO: 1) from nucleotide 374 to nucleotide 1291.
12. A vector comprising the polynucleotide of claim 2.
13. A host cell comprising the vector of claim 12.
14. A method of producing a polypeptide comprising expressing from the host cell of claim 13 the polypeptide encoded by the polynucleotide.
15. The method of claim 14 wherein the polypeptide comprises amino acid 1 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2).
16. The method of claim 14 wherein the polypeptide comprises amino acid 21 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2).
17. The method of claim 14 wherein the polypeptide comprises amino acid 27 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2).
18. A method of producing a polypeptide wherein the polypeptide comprises the amino acid sequence shown in FIG. 2 (SEQ ID NO: 2), the method comprising the steps of:
(a) culturing the host cell of claim 13 under conditions whereby the polypeptide is expressed; and
(b) recovering the polypeptide from the culture.
19. A method for producing a cell which expresses a polypeptide comprising genetically engineering the cell with the vector of claim 12.
20. A polypeptide comprising a member selected from the group consisting of:
(a) a polypeptide, or a biologically or immunologically active fragment thereof, comprising the amino acid sequence as set forth in FIG. 2 (SEQ ID NO: 2);
(b) a polypeptide comprising amino acid 21 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2);
(c) a polypeptide comprising amino acid 27 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2);
(d) a polypeptide comprising amino acid 28 to amino acid 46 as set forth in FIG. 2 (SEQ ID NO: 2);
(e) a polypeptide comprising amino acid 77 to amino acid 91 as set forth in FIG. 2 (SEQ ID NO: 2); and
(f) a polypeptide which is at least 70% identical to the polypeptide of (a), (b), (c), (d), or (e).
21. The polypeptide of claim 20 wherein the polypeptide comprises amino acids 1 to 331 as set forth in FIG. 2 (SEQ ID NO: 2).
22. The polypeptide of claim 20 wherein the polypeptide comprises amino acids 21 to 331 as set forth in FIG. 2 (SEQ ID NO: 2).
23. The polypeptide of claim 20 wherein the polypeptide comprises amino acids 27 to 331 as set forth in FIG. 2 (SEQ ID NO: 2).
24. An isolated antibody, or antibody fragment, which specifically binds to a polypeptide comprising a member selected from the group consisting of:
(a) a polypeptide, or a biologically or immunologically active fragment thereof, comprising the amino acid sequence as set forth in FIG. 2 (SEQ ID NO: 2);
(b) a polypeptide comprising amino acid 28 to amino acid 46 as set forth in FIG. 2 (SEQ ID NO: 2);
(c) a polypeptide comprising amino acid 77 to amino acid 91 as set forth in FIG. 2 (SEQ ID NO: 2);
(d) a polypeptide comprising amino acid 188 to amino acid 210 as set forth in FIG. 2 (SEQ ID NO: 2);
(e) a polypeptide comprising amino acid 263 to amino acid 274 as set forth in FIG. 2 (SEQ ID NO: 2); and
(f) a polypeptide which is at least 70% identical to the polypeptide of (a), (b), (c), (d), or (e).
25. The antibody of claim 24, wherein the antibody specifically binds to the amino acid sequence PLGGESICSAGAPAKYSIT (SEQ ID NO: 8).
26. The antibody of claim 24, wherein the antibody specifically binds to the amino acid sequence HSSDYSMWRKNQYVS (SEQ ID NO: 10).
27. The antibody of claim 24, wherein the antibody specifically binds to the amino acid sequence DAGTDSGFTFSSPNFATIPQDTV (SEQ ID NO: 11).
28. The antibody of claim 24, wherein the antibody specifically binds to the amino acid sequence NEIVDSASVPET (SEQ ID NO: 12).
29. The antibody of claim 24, wherein the antibody is a polyclonal antibody.
30. The antibody of claim 24, wherein the antibody is a monoclonal antibody.
31. An immunoconjugate comprising an isolated antibody, or antibody fragment, which specifically binds to a polypeptide comprising a member selected from the group consisting of:
(a) a polypeptide, or a biologically or immunologically active fragment thereof, comprising the amino acid sequence as set forth in FIG. 2 (SEQ ID NO: 2);
(b) a polypeptide comprising amino acid 21 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2);
(c) a polypeptide comprising amino acid 27 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2);
(d) a polypeptide comprising amino acid 28 to amino acid 46 as set forth in FIG. 2 (SEQ ID NO: 2);
(e) a polypeptide comprising amino acid 77 to amino acid 91 as set forth in FIG. 2 (SEQ ID NO: 2);
(f) a polypeptide comprising amino acid 188 to amino acid 210 as set forth in FIG. 2 (SEQ ID NO: 2);
(g) a polypeptide comprising amino acid 263 to amino acid 274 as set forth in FIG. 2 (SEQ ID NO: 2); and
(h) a polypeptide which is at least 70% identical to the polypeptide of (a), (b), (c), (d), (e), (f), or (g)
conjugated to a therapeutic agent.
32. The immunoconjugate of claim 31, wherein the therapeutic agent is a cytotoxic agent.
33. The immunoconjugate of claim 32, wherein the cytotoxic agent is selected from the group consisting of ricin, doxorubicin, daunorubicin, taxol, ethidium bromide, mitomycin, etoposied, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin D, diphteria toxin, Pseudomonas exotoxin (PE) A, PE40, ricin, abrin, glucodorticoid and radioisotopes.
34. The immunoconjugate of claim 31, wherein the antibody fragments are selected from the group consisting of Fv, F(ab) and F(ab)2 fragments.
35. A method for selectively destroying a cell expressing the polypeptide of FIG. 2 (SEQ ID NO: 2) comprising reacting the immunoconjugate of claim 31 with the cell so that the therapeutic agent of the immunoconjugate can destroy the cell.
36. A method of treating a disease-state in a human patient which disease-state is associated with expression of RG1 and wherein the method comprises administering to the patient a therapeutically effective amount of the immunoconjugate of claim 31.
37. A method of treating a disease-state in a human patient which disease-state is associated with inappropriate expression of RG1 and wherein the patient is in need of decreased levels of a polypeptide comprising a member selected from the group consisting of:
(a) a polypeptide, or a biologically or immunologically active fragment thereof, comprising the amino acid sequence as set forth in FIG. 2 (SEQ ID NO: 2); and
(b) a polypeptide which is at least 70% identical to the polypeptide of (a) and wherein the method comprises administering to the patient a therapeutically effective amount of a ribozyme which specifically cleaves RNA encoding the polypeptide.
38. A method of treating a disease-state in a human patient which disease-state is associated with inappropriate expression of RG1 and wherein the patient is in need of decreased levels of a polypeptide having the amino acid sequence as set forth in FIG. 2 (SEQ ID NO: 2); wherein the method comprises administering to the patient a therapeutically effective amount of a polynucleotide which is complementary to a polynucleotide encoding the polypeptide or a portion thereof.
39. A diagnostic method wherein the method comprises analyzing a sample derived from a host for the presence of a polypeptide comprising a member selected from the group consisting of:
(a) a polypeptide, or a biologically or immunologically active fragment thereof, comprising the amino acid sequence as set forth in FIG. 2 (SEQ ID NO: 2);
(b) a polypeptide comprising amino acid 21 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2);
(c) a polypeptide comprising amino acid 27 to amino acid 331 as set forth in FIG. 2 (SEQ ID NO: 2);
(d) a polypeptide comprising amino acid 28 to amino acid 46 as set forth in FIG. 2 (SEQ ID NO: 2);
(e) a polypeptide comprising amino acid 77 to amino acid 91 as set forth in FIG. 2 (SEQ ID NO: 2);
(e) a polypeptide comprising amino acid 188 to amino acid 210 as set forth in FIG. 2 (SEQ ID NO: 2);
(g) a potypeptide comprising amino acid 263 to amino acid 274 as set forth in FIG. 2 (SEQ ID NO: 2); and
(h) a polypeptide which is at least 70% identical to the polypeptide of (a), (b), (c), (d), (e), (f), or (g).
40. The method of claim 39, wherein analyzing comprises contacting the sample with the antibody or antibody fragment of claim 24, which specifically binds to the polypeptide and detecting binding of the antibody to the polypeptide in the sample.
41. A diagnostic method wherein the method comprises analyzing for the presence of a polynucleotide comprising a polynucleotide which is at least 70% identical to a member selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide, or a biologically or immunologically active fragment thereof, comprising the amino acid sequence set forth in FIG. 2 (SEQ ID NO: 2); and
(b) polynucleotide which is complementary to the polynucleotide of (a).
42. A method for diagnosing in a subject a metastasis associated with the polypeptide of FIG. 2 (SEQ ID NO: 2) comprising:
(a) obtaining from the subject a tissue andor fluid sample;
(b) contacting the sample with the antibody of claim 24; and
(c) detecting the binding of the antibody with the polypeptide in the sample.
43. The method of claim 42, wherein the antibody is labeled so as to directly or indirectly produce a detectable signal with a compound selected from the group consisting of a radiolabel, an enzyme, a chromophore and a fluorescer.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A screed die box comprising:
a screed die bucket;
a screed die box gate slidably connected at the bottom of the screed die bucket;
a screed die box lever rotatably connected to the screed die bucket and the screed die box gate for sliding the screed die box gate between an open position and a closed position;
a screed plate slidably connected along an aft side of the screed die bucket, the screed plate including a positioning aperture; and
a positioning member that is located in the screed die bucket that engages the screed plate, the positioning member sliding the screed plate as the positioning member is rotated.
2. The screed die box of claim 1, wherein the positioning member is substantially parallel to the aft side of the screed die bucket.
3. The screed die box of claim 1, and further comprising:
a screed die box anchor attached to a forward side of the screed die bucket.
4. The screed die box of claim 1, wherein the screed plate comprises:
a first portion that extends substantially parallel to the aft side of the screed die bucket; and
a second portion that extends substantially perpendicularly to the first portion, the second portion including the positioning aperture.
5. The screed die box of claim 4, and further comprising:
a retaining slot in the first portion of the screed plate; and
a retaining screw positioned in the retaining slot and located in the screed die bucket.
6. The screed die box of claim 1, wherein the positioning member comprises:
a screw shaft;
a screw head at one end of the screw shaft; and
a screw shoulder positioned along the length of the screw shaft that is spaced apart from the screw head;
wherein the positioning member engages the screed plate by having the screed plate positioned between the screw head and the screw shoulder.
7. The screed die box of claim 1, wherein the screed die box gate contacts the aft side of the screed die bucket when the screed die box gate is in the closed position.
8. The screed die box of claim 1, wherein a thermoplastic material enters into the screed die box through the top of the screed die box and exits out of the bottom of the screed die box when the screed die box gate is in the open position.
9. The screed die box of claim 1, wherein the screed die box gate slides generally vertically and the screed plate slides generally horizontally.
10. A mobile applicator comprising:
a frame;
a plurality of wheels rotatably connected to the frame;
a kettle attached to the frame for holding a thermoplastic material; and
a screed die box positioned to receive thermoplastic material from the kettle, the screed die box comprising:
a screed die bucket;
a screed plate at the rear of the screed die bucket; and
a positioning member that is located in the screed die bucket that engages the screed plate, the positioning member sliding the screed plate as the positioning member is rotated.
11. The mobile applicator of claim 10, wherein the screed die box further comprises:
a screed die box gate slidably connected at the bottom of the screed die bucket; and
a screed die box lever rotatably connected to the screed die bucket and the screed die box gate for sliding the screed die box gate between an open position and a closed position.
12. The mobile applicator of claim 11, wherein the screed die box gate contacts the aft side of the screed die bucket when the screed die box gate is in the closed position.
13. The mobile applicator of claim 11, wherein thermoplastic material enters into the screed die box through the top of the screed die box and exits out of the bottom of the screed die box when the screed die box gate is in the open position.
14. The mobile applicator of claim 11, and further comprising:
a screed actuator lever attached to the screed die box lever for rotating the screed die box lever.
15. The mobile applicator of claim 10, wherein the positioning member is substantially parallel to the aft side of the screed die bucket.
16. The mobile applicator of claim 10, and further comprising:
a screed die box anchor attached to a forward side of the screed die bucket.
17. The mobile applicator of claim 10, wherein the screed plate comprises:
a first portion that extends substantially parallel to the aft side of the screed die bucket; and
a second portion that extends substantially perpendicularly to the first portion, the second portion including the positioning aperture.
18. The mobile applicator of claim 15, and further comprising:
a retaining slot in the first portion of the screed plate; and
a retaining screw positioned in the retaining slot and located in the screed die bucket.
19. The mobile applicator of claim 10, wherein the positioning member comprises:
a screw shaft;
a screw head at one end of the screw shaft; and
a screw shoulder along the length of the screw shaft that is spaced apart from the screw head;
wherein the positioning member engages the screed plate by having the screed plate positioned between the screw head and the screw shoulder.
20. The mobile applicator of claim 10, and further comprising:
a chute connected to a side of the kettle near a bottom of the kettle;
wherein the screed die box is positioned beneath the chute.