Claims
- 1. An isolated polynucleotide encoding a zpep14 polypeptide comprising a sequence of amino acid residues that is at least 90% identical to an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 92 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 137 (Ile); (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 171 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 137 (Ile); (e) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 171 (Gly); (f) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 171 (Gly); (g) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 174 (Ile) to amino acid number 188 (Asn); (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 188 (Asn); (i) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 188 (Asn); (j) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); and (k) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 188 (Asn), wherein the amino acid percent identity is determined using a FASTA program with ktup=1, gap opening penalty=10, gap extension penalty=1, and substitution matrix=BLOSUM62, with other parameters set as default.
- 2. An isolated polynucleotide according to claim 1, wherein the polynucleotide is selected from the group consisting of:
(a) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 53 to nucleotide 280; (b) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 290 to nucleotide 415; (c) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 422 to nucleotide 517; (d) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 53 to nucleotide 415; (e) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 53 to nucleotide 517; (f) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 290 to nucleotide 517; (g) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 524 to nucleotide 568; (h) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 422 to nucleotide 568; (i) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 290 to nucleotide 568; (j) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 53 to nucleotide 568; (k) a polynucleotide sequence as shown in SEQ ID NO:1 from nucleotide 5 to nucleotide 568; and (l) a polynucleotide sequence complementary to (a) through (k).
- 3. An isolated polynucleotide sequence according to claim 1, wherein the polynucleotide comprises nucleotide 1 to nucleotide 564 of SEQ ID NO:3.
- 4. An isolated polynucleotide according to claim 1, wherein the zpep14 polypeptide comprises a sequence of amino acid residues selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 92 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 137 (Ile); (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 171 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 137 (Ile); (e) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 171 (Gly); (f) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 171 (Gly); (g) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 174 (Ile) to amino acid number 188 (Asn); (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 188 (Asn); (i) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 188 (Asn); (j) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); and (k) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 188 (Asn).
- 5. An isolated polynucleotide according to claim 4, wherein the zpep14 polypeptide consists of a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 137 (lIe).
- 6. An expression vector comprising the following operably linked elements:
a transcription promoter; a DNA segment encoding a zpep14 polypeptide that is at least 90% identical to an amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); and a transcription terminator.
- 7. An expression vector according to claim 6, further comprising a secretory signal sequence operably linked to the DNA segment.
- 8. A cultured cell into which has been introduced an expression vector according to claim 6, wherein the cell expresses a polypeptide encoded by the DNA segment.
- 9. A DNA construct encoding a fusion protein, the DNA construct comprising:
a first DNA segment encoding a polypeptide that is at least 90% identical to a sequence of amino acid residues selected from the group consisting of:
(a) the amino acid sequence of SEQ ID NO:2 from residue number 1 (Met) to residue number 16 (Ala)); (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 92 (Cys); (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 137 (Ile); (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 171 (Gly); (e) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 137 (Ile); (f) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 171 (Gly); (g) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 171 (Gly); (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 174 (Ile) to amino acid number 188 (Asn); (i) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 188 (Asn); (j) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 188 (Asn); (k) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); and (l) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 188 (Asn). at least one other DNA segment encoding an additional polypeptide, wherein the first and other DNA segments are connected in-frame; and encode the fusion protein.
- 10. A fusion protein produced by a method comprising:
culturing a host cell into which has been introduced a vector comprising the following operably linked elements:
(a) a transcriptional promoter; (b) a DNA construct encoding a fusion protein according to claim 9; and (c) a transcriptional terminator; and recovering the protein encoded by the DNA segment.
- 11. An isolated polypeptide comprising a sequence of amino acid residues that is at least 90% identical to an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 92 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 137 (Ile); (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 171 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 137 (Ile); (e) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 171 (Gly); (f) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 171 (Gly); (g) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 174 (Ile) to amino acid number 188 (Asn); (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 188 (Asn); (i) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 188 (Asn); (j) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); and (k) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 188 (Asn), wherein the amino acid percent identity is determined using a FASTA program with ktup=1, gap opening penalty=10, gap extension penalty=1, and substitution matrix=BLOSUM62, with other parameters set as default.
- 12. An isolated polypeptide according to claim 11, wherein the polypeptide comprising a sequence of amino acid residues that is selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 92 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 137 (Ile); (c) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 171 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 137 (Ile); (e) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 171 (Gly); (f) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 171 (Gly); (g) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 174 (Ile) to amino acid number 188 (Asn); (h) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 140 (Gln) to amino acid number 188 (Asn); (i) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 188 (Asn); (j) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); and (k) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met) to amino acid number 188 (Asn).
- 13. An isolated polypeptide according to claim 12, wherein the sequence of amino acid residues is as shown in SEQ ID NO:2 from amino acid number 96 (Asn) to amino acid number 137 (Ile).
- 14. A method of producing a zpep14 polypeptide comprising:
culturing a cell according to claim 8; and isolating the zpep14 polypeptide produced by the cell.
- 15. A method of detecting, in a test sample, the presence of a modulator of zpep14 protein activity, comprising:
transfecting a zpep14-responsive cell, with a reporter gene construct that is responsive to a zpep14-stimulated cellular pathway; and producing a zpep14 polypeptide by the method of claim 14; and adding the zpep14 polypeptide to the cell, in the presence and absence of a test sample; and comparing levels of response to the zpep14 polypeptide, in the presence and absence of the test sample, by a biological or biochemical assay; and determining from the comparison, the presence of the modulator of zpep14 activity in the test sample.
- 16. A method of producing an antibody to zpep14 polypeptide comprising the following steps in order:
inoculating an animal with a polypeptide selected from the group consisting of:
(a) a polypeptide consisting of 9 to 172 amino acids, wherein the polypeptide is at least 90% identical to a contiguous sequence of amino acids in SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); (b) a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 from amino acid number 17 (Arg) to amino acid number 188 (Asn); (c) a polypeptide according to claim 11;(d) a polypeptide consisting of amino acid number 90 (Asn) to amino acid number 95 (Arg) of SEQ ID NO:2; (e) a polypeptide consisting amino acid number 128 (Glu) to amino acid number 133 (Glu) of SEQ ID NO:2; (f) a polypeptide consisting of amino acid number 167 (Glu) to amino acid number 172 (Lys) of SEQ ID NO:2; (g) a polypeptide consisting of amino acid number 175 (Ile) to amino acid number 180 (Lys) of SEQ ID NO:2; and (h) a polypeptide consisting of amino acid number 176 (Glu) to amino acid number 181 (Arg) of SEQ ID NO:2; and and wherein the polypeptide elicits an immune response in the animal to produce the antibody; and isolating the antibody from the animal.
- 17. An antibody produced by the method of claim 16, which binds to a zpep14 polypeptide.
- 18. The antibody of claim 17, wherein the antibody is a monoclonal antibody.
- 19. An antibody which binds to a polypeptide of claim 11.
- 20. An isolated polynucleotide encoding a zpep14 polypeptide comprising a sequence of amino acid residues that is at least 90% identical to an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 91 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 170 (Gly); (c) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 95 (Asn) to amino acid number 170 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 173 (Ile) to amino acid number 187 (Asn); (e) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 187 (Asn); and (f) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 1 (Met) to amino acid number 187 (Asn), wherein the amino acid percent identity is determined using a FASTA program with ktup=1, gap opening penalty=10, gap extension penalty=1, and substitution matrix=BLOSUM62, with other parameters set as default.
- 21. An isolated polynucleotide according to claim 20, wherein the polynucleotide is selected from the group consisting of:
(a) a polynucleotide sequence as shown in SEQ ID NO:17 from nucleotide 96 to nucleotide 320; (b) a polynucleotide sequence as shown in SEQ ID NO:17 from nucleotide 96 to nucleotide 557; (c) a polynucleotide sequence as shown in SEQ ID NO:17 from nucleotide 330 to nucleotide 557; (d) a polynucleotide sequence as shown in SEQ ID NO:17 from nucleotide 564 to nucleotide 608; (e) a polynucleotide sequence as shown in SEQ ID NO:17 from nucleotide 96 to nucleotide 608; (f) a polynucleotide sequence as shown in SEQ ID NO:17 from nucleotide 48 to nucleotide 608; and (g) a polynucleotide sequence complementary to (a) through (k).
- 22. An isolated polynucleotide according to claim 20, wherein the zpep14 polypeptide comprises a sequence of amino acid residues selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 91 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 170 (Gly); (c) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 95 (Asn) to amino acid number 170 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 173 (Ile) to amino acid number 187 (Asn); (e) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 187 (Asn); and (f) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 1 (Met) to amino acid number 187 (Asn).
- 23. An expression vector comprising the following operably linked elements:
a transcription promoter; a DNA segment encoding a zpep14 polypeptide that is at least 90% identical to an amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 187 (Asn); and a transcription terminator.
- 24. An expression vector according to claim 23, further comprising a secretory signal sequence operably linked to the DNA segment.
- 25. A cultured cell into which has been introduced an expression vector according to claim 23, wherein the cell expresses a polypeptide encoded by the DNA segment.
- 26. An isolated polypeptide comprising a sequence of amino acid residues that is at least 90% identical to an amino acid sequence selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 91 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 170 (Gly); (c) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 95 (Asn) to amino acid number 170 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 173 (Ile) to amino acid number 187 (Asn); (e) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 187 (Asn); and (f) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 1 (Met) to amino acid number 187 (Asn), wherein the amino acid percent identity is determined using a FASTA program with ktup=1, gap opening penalty=10, gap extension penalty=1, and substitution matrix=BLOSUM62, with other parameters set as default.
- 27. An isolated polypeptide according to claim 26, wherein the polypeptide comprises a sequence of amino acid residues that is selected from the group consisting of:
(a) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 91 (Cys); (b) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 170 (Gly); (c) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 95 (Asn) to amino acid number 170 (Gly); (d) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 173 (Ile) to amino acid number 187 (Asn); (e) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 17 (Gly) to amino acid number 187 (Asn); and (f) the amino acid sequence as shown in SEQ ID NO:18 from amino acid number 1 (Met) to amino acid number 187 (Asn).
- 28. A method of producing a zpep14 polypeptide comprising:
culturing a cell according to claim 25; and isolating the zpep14 polypeptide produced by the cell.
- 29. A method of producing an antibody to a zpep14 polypeptide comprising the following steps in order:
inoculating an animal with a polypeptide selected from the group consisting of:
(a) a polypeptide according to claim 26;(b) a polypeptide consisting of the amino acid sequence of SEQ ID NO:18 from amino acid number 17 (Gly) to 91 (Cys); (c) a polypeptide consisting of the amino acid sequence of SEQ ID NO:18 from amino acid number 17 (Gly) to 170 (Gly); (d) a polypeptide consisting of the amino acid sequence of SEQ ID NO:18 from amino acid number 95 (Asn) to 170 (Gly); (e) a polypeptide consisting of the amino acid sequence of SEQ ID NO:18 from amino acid number 173 (Ile) to 187 (Asn); (f) a polypeptide consisting of the amino acid sequence of SEQ ID NO:18 from amino acid number 17 (Gly) to 187 (Asn); and (g) a polypeptide consisting of a hydrophilic peptide predicted from a murine zpep14 hydrophobicity plot using a Hopp/Woods hydrophilicity profile based on a sliding six-residue window, with buried G, S, and T residues and exposed H, Y, and W residues ignored; and and wherein the polypeptide elicits an immune response in the animal to produce the antibody; and isolating the antibody from the animal.
- 30. An antibody produced by the method of claim 29, which binds to a zpep14 polypeptide.
- 31. The antibody of claim 30, wherein the antibody is a monoclonal antibody.
- 32. An antibody which binds to a polypeptide of claim 26.
REFERENCE TO RELATED APPLICATIONS
[0001] This application is related to Provisional Application 60/119,173, filed on Feb. 8, 1999. This application is also related to Provisional Application 60/151,035, filed on Aug. 27, 1999. Under 35 U.S.C. §119(e)(1), this application claims benefit of said Provisional Applications.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60119173 |
Feb 1999 |
US |
|
60151035 |
Aug 1999 |
US |