Claims
- 1. An isolated protein comprising a polypeptide that is at least 80% identical to a polypeptide selected from the group consisting of:
a) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 65 of SEQ ID NO:2; b) a polypeptide having the sequence of amino acid residue 19 to amino acid residue 65 of SEQ ID NO:2; c) a polypeptide having the sequence of amino acid residue 21 to amino acid residue 65 of SEQ ID NO:2; d) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 67 of SEQ ID NO:10; e) a polypeptide having the sequence of amino acid residue 21 to amino acid residue 67 of SEQ ID NO:10; and f) a polypeptide having the sequence of amino acid residue 23 to amino acid residue 67 of SEQ ID NO:2; wherein said polypeptide has cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NOs:2 or 10.
- 2. An isolated protein of claim 1, 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.
- 3. An isolated protein of claim 1, wherein said protein comprises a polypeptide having the sequence selected from the group consisting of:
a) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 67 of SEQ ID NO:10; b) a polypeptide having the sequence of amino acid residue 21 to amino acid residue 67 of SEQ ID NO:10; and c) a polypeptide having the sequence of amino acid residue 23 to amino acid residue 67 of SEQ ID NO:10.
- 4. A polypeptide selected from the group consisting of:
a) amino acid residue 30 to amino acid residue 63 of SEQ ID NO:2; b) amino acid residue 31 to amino acid residue 63 of SEQ ID NO:2; c) amino acid residue 30 to amino acid residue 64 of SEQ ID NO:2; d) amino acid residue 31 to amino acid residue 64 of SEQ ID NO:2; and e) a polypeptide chosen from SEQ ID NOs:14-72.
- 5. A pharmaceutical composition comprising a polypeptide selected from the group consisting of:
a) a protein according to claim 1;b) amino acid residue 30 to amino acid residue 63 of SEQ ID NO:2; c) amino acid residue 31 to amino acid residue 63 of SEQ ID NO:2; d) amino acid residue 30 to amino acid residue 64 of SEQ ID NO:2; e) amino acid residue 31 to amino acid residue 64 of SEQ ID NO:2; and f) a polypeptide chosen from SEQ ID NOs:14-72; in combination with a pharmaceutically acceptable vehicle.
- 6. An antibody that specifically binds to a protein of claim 1.
- 7. An anti-idiotypic antibody of an antibody which specifically binds to a protein of claim 1.
- 8. An isolated polynucleotide molecule encoding a protein, said polynucleotide molecule consisting of a coding strand and a complementary non-coding strand, wherein said polynucleotide molecule encodes a polypeptide that is at least 80% identical to the amino acid sequence to a polypeptide selected from the group consisting of:
a) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 65 of SEQ ID NO:2; b) a polypeptide having the sequence of amino acid residue 19 to amino acid residue 65 of SEQ ID NO:2; c) a polypeptide having the sequence of amino acid residue 21 to amino acid residue 65 of SEQ ID NO:2; d) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 67 of SEQ ID NO:10; e) a polypeptide having the sequence of amino acid residue 21 to amino acid residue 67 of SEQ ID NO:10; and f) a polypeptide having the sequence of amino acid residue 23 to amino acid residue 67 of SEQ ID NO:2; wherein said polypeptide has cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NOs:2 or 10.
- 9. An isolated polynucleotide molecule according to claim 8, 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.
- 10. An isolated polynucleotide molecule according to claim 8, wherein said polynucleotide molecule remains hybridized following stringent wash conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:9, or the complement of SEQ ID NO:9.
- 11. An isolated polynucleotide molecule encoding a protein having cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NO:10, said polynucleotide molecule consisting of a coding strand and a complementary non-coding strand, wherein said polynucleotide comprises a nucleotide sequence that is at least 80% identical to the sequence of a polynucleotide selected from the group consisting of:
a) a polynucleotide as shown in SEQ ID NO:9 from nucleotide 220 to nucleotide 420; b) a polynucleotide as shown in SEQ ID NO:9 from nucleotide 280 to nucleotide 420; and c) a polynucleotide as shown in SEQ ID NO:9 from nucleotide 286 to nucleotide 420.
- 12. An isolated polynucleotide molecule encoding a protein having cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NO:10, said polynucleotide molecule consisting of a coding strand and a complementary non-coding strand, wherein said polynucleotide comprises a nucleotide sequence as shown in SEQ ID NO:11.
- 13. An isolated polynucleotide molecule encoding a polypeptide selected from the group consisting of:
a) amino acid residue 30 to amino acid residue 63 of SEQ ID NO:2; b) amino acid residue 31 to amino acid residue 63 of SEQ ID NO:2; c) amino acid residue 30 to amino acid residue 64 of SEQ ID NO:2; d) amino acid residue 31 to amino acid residue 64 of SEQ ID NO:2; and e) a polypeptide chosen from SEQ ID NOs:14-72.
- 14. An isolated polynucleotide molecule selected from the group consisting of:
a) nucleotide 88 to nucleotide 189 of SEQ ID NO:1; b) nucleotide 88 to nucleotide 192 of SEQ ID NO:1; c) nucleotide 91 to nucleotide 189 of SEQ ID NO:1; d) nucleotide 91 to nucleotide 192 of SEQ ID NO:1; e) nucleotide 88 to nucleotide 189 of SEQ ID NO:4; f) nucleotide 88 to nucleotide 192 of SEQ ID NO:4; g) nucleotide 91 to nucleotide 189 of SEQ ID NO:4 and h) nucleotide 91 to nucleotide 192 of SEQ ID NO:4.
- 15. An expression vector comprising the following operably linked elements:
a transcription promoter; a DNA segment encoding a polypeptide selected from the group consisting of:
a) a protein of claim 1;b) amino acid residue 30 to amino acid residue 63 of SEQ ID NO:2; c) amino acid residue 31 to amino acid residue 63 of SEQ ID NO:2; d) amino acid residue 30 to amino acid residue 64 of SEQ ID NO:2; e) amino acid residue 31 to amino acid residue 64 of SEQ ID NO:2; and f) a polypeptide chosen from SEQ ID NOs:14-72; and a transcription terminator.
- 16. An expression vector according to claim 15, wherein said DNA segment further encodes a secretory signal sequence operably linked to said protein.
- 17. An expression vector according the claim 16, wherein said secretory signal sequence is selected from the group consisting of:
a) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 18 of SEQ ID NO:2; b) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 20 of SEQ ID NO:2; c) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 20 of SEQ ID NO:10; and d) a polypeptide having the sequence of amino acid residue 1 to amino acid residue 22 of SEQ ID NO:10.
- 18. A cultured cell into which has been introduced an expression vector according to claim 15;wherein said cell expresses said polypeptide encoded by said DNA segment.
- 19. A method of producing a protein comprising: culturing a cell into which has been introduced an expression vector according to claim 15;whereby said cell expresses said polypeptide encoded by said DNA segment; and recovering said expressed polypeptide.
- 20. An oligonucleotide probe or primer comprising at least 14 contiguous nucleotides of a polynucleotide of SEQ ID NO:11 or a sequence complementary to SEQ ID NO:11.
- 21. A method of treating a microbial-related disease comprising administering to a mammal a therapeutically effective amount of a polypeptide selected from the group consisting of:
a) a polypeptide of SEQ ID NO:2; b) a polypeptide of SEQ ID NO:10; c) a polypeptide chosen from SEQ ID NOs:14-72; d) amino acid residue 30 to amino acid residue 63 of SEQ ID NO:2; e) amino acid residue 31 to amino acid residue 63 of SEQ ID NO:2; f) amino acid residue 30 to amino acid residue 64 of SEQ ID NO:2; g) amino acid residue 31 to amino acid residue 64 of SEQ ID NO:2; h) amino acid residue 20 to amino acid residue 67 of SEQ ID NO:10 and i) amino acid residue 22 to amino acid residue 67 of SEQ ID NO:10; whereby said polypeptide ameliorates said disease.
- 22. A method of claim 21, wherein said microbial-related disease is associated with the eye.
- 23. A method of claim 22, wherein said microbial-related disease is conjunctivitis.
- 24. A method of claim 21, wherein said microbial-related disease is associated with the ear.
- 25. A method of contraception comprising administering to a mammal a therapeutically effective amount of a polypeptide selected from the group consisting of:
a) a polypeptide of SEQ ID NO:2; b) a polypeptide of SEQ ID NO:10; c) a polypeptide chosen from SEQ ID NOs:14-72; d) amino acid residue 30 to amino acid residue 63 of SEQ ID NO:2; e) amino acid residue 31 to amino acid residue 63 of SEQ ID NO:2; f) amino acid residue 30 to amino acid residue 64 of SEQ ID NO:2; g) amino acid residue 31 to amino acid residue 64 of SEQ ID NO:2; h) amino acid residue 20 to amino acid residue 67 of SEQ ID NO:10 and i) amino acid residue 22 to amino acid residue 67 of SEQ ID NO:10.
- 26. An isolated polynucleotide molecule encoding a protein, wherein the encoded protein comprises an amino acid sequence having at least 95 percent sequence identity with amino acid residues 31 to 63 of SEQ ID NO:2, and wherein the encoded protein has antimicrobial activity.
- 27. The isolated polynucleotide molecule of claim 26 wherein the encoded protein comprises amino acid residues selected from the group of 30 to 63 of SEQ ID NO:2, 30 to 64 of SEQ ID NO:2, 31 to 64 of SEQ ID NO:2, 21 to 65 of SEQ ID NO:2, 19 to 65 of SEQ ID NO:2, and SEQ ID NO:2.
- 28. The isolated polynucleotide molecule of claim 26 wherein the encoded protein has cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NO:2.
- 29. The isolated polynucleotide molecule of claim 26 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.
- 30. The isolated polynucleotide molecule of claim 26 wherein the differences between the encoded protein and amino acid residues 31 to 63 of SEQ ID NO:2 are due to conservative amino acid substitutions.
- 31. The isolated polynucleotide molecule of claim 26 wherein the encoded protein has antimicrobial activity to at least one microbe selected from the group of bacteria, anaerobic organisms, protozoans, fungi, and viruses.
- 32. The isolated polynucleotide molecule of claim 26 wherein the encoded protein has antimicrobial activity to bacteria.
- 33. The isolated polynucleotide molecule of claim 32 wherein the bacteria is gram positive.
- 34. The isolated polynucleotide molecule of claim 32 wherein the bacteria is gram negative.
- 35. An isolated polynucleotide molecule encoding a protein, wherein the encoded protein consists of an amino acid sequence having at least 95 percent sequence identity with amino acid residues 31 to 63 of SEQ ID NO:2, and wherein the encoded protein has antimicrobial activity.
- 36. The isolated polynucleotide molecule of claim 35 wherein the encoded protein consists of amino acid residues selected from the group of 30 to 63 of SEQ ID NO:2, 30 to 64 of SEQ ID NO:2, 31 to 64 of SEQ ID NO:2, 21 to 65 of SEQ ID NO:2, 19 to 65 of SEQ ID NO:2, and SEQ ID NO:2.
- 37. The isolated polynucleotide molecule of claim 35 wherein the encoded protein has cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NO:2.
- 38. The isolated polynucleotide molecule of claim 35 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.
- 39. The isolated polynucleotide molecule of claim 35 wherein the differences between the encoded protein and amino acid residues 31 to 63 of SEQ ID NO:2 are due to conservative amino acid substitutions.
- 40. The isolated polynucleotide molecule of claim 35 wherein the encoded protein has antimicrobial activity to at least one microbe selected from the group of bacteria, anaerobic organisms, protozoans, fungi, and viruses.
- 41. The isolated polynucleotide molecule of claim 35 wherein the encoded protein has antimicrobial activity to bacteria.
- 42. The isolated polynucleotide molecule of claim 41 wherein the bacteria is gram positive.
- 43. The isolated polynucleotide molecule of claim 41 wherein the bacteria is gram negative.
- 44. An isolated polynucleotide molecule encoding a protein, wherein the encoded protein comprises an amino acid sequence having at least 95 percent sequence identity with amino acid residues 23 to 67 of SEQ ID NO:10, and wherein the encoded protein has antimicrobial activity.
- 45. The isolated polynucleotide molecule of claim 44 wherein the encoded protein comprises amino acid residues selected from the group of 22 to 67 of SEQ ID NO:10, 21 to 67 of SEQ ID NO:10, 20 to 67 of SEQ ID NO:10, and SEQ ID NO:10.
- 46. The isolated polynucleotide molecule of claim 44 wherein the encoded protein has cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NO:10.
- 47. The isolated polynucleotide molecule of claim 44 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.
- 48. The isolated polynucleotide molecule of claim 44 wherein the differences between the encoded protein and amino acid residues 23 to 67 of SEQ ID NO:10 are due to conservative amino acid substitutions.
- 49. The isolated polynucleotide molecule of claim 44 wherein the encoded protein has antimicrobial activity to at least one microbe selected from the group of bacteria, anaerobic organisms, protozoans, fungi, and viruses.
- 50. The isolated polynucleotide molecule of claim 44 wherein the encoded protein has antimicrobial activity to bacteria.
- 51. The isolated polynucleotide molecule of claim 50 wherein the bacteria is gram positive.
- 52. The isolated polynucleotide molecule of claim 50 wherein the bacteria is gram negative.
- 53. An isolated polynucleotide molecule that encoding a protein, wherein the encoded protein consists of an amino acid sequence having at least 95 percent sequence identity with amino acid residues 23 to 67 of SEQ ID NO:10, and wherein the encoded protein has antimicrobial activity.
- 54. The isolated polynucleotide molecule of claim 53 wherein the encoded protein consists of amino acid residues 22 to 67 of SEQ ID NO:10, 21 to 67 of SEQ ID NO:10, 20 to 67 of SEQ ID NO:10, and SEQ ID NO:10.
- 55. The isolated polynucleotide molecule of claim 53 wherein the encoded protein has cysteine residues corresponding to amino acid residues 33, 40, 45, 55, 62 and 63 of SEQ ID NO:10.
- 56. The isolated polynucleotide molecule of claim 53 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.
- 57. The isolated polynucleotide molecule of claim 53 wherein the differences between the encoded protein and amino acid residues 23 to 67 of SEQ ID NO:10 are due to conservative amino acid substitutions.
- 58. The isolated polynucleotide molecule of claim 53 wherein the encoded protein has antimicrobial activity to at least one microbe selected from the group of bacteria, anaerobic organisms, protozoans, fungi, and viruses.
- 59. The isolated polynucleotide molecule of claim 53 wherein the encoded protein has antimicrobial activity to bacteria.
- 60. The isolated polynucleotide molecule of claim 59 wherein the bacteria is gram positive.
- 61. The isolated polynucleotide molecule of claim 59 wherein the bacteria is gram negative.
- 62. An expression vector comprising the following operably linked element:
a transcription promoter; a DNA segment comprising the isolated polynucleotide molecule encoding a protein of claim 26; and a transcription promoter.
- 63. The expression vector of claim 62 wherein the DNA segment further encodes a secretory signal sequence operably linked to the protein.
- 64. An expression vector comprising the following operably linked element:
a transcription promoter; a DNA segment comprising the isolated polynucleotide molecule encoding a protein of claim 44; and a transcription promoter.
- 65. The expression vector of claim 64 wherein the DNA segment further encodes a secretory signal sequence operably linked to the protein.
- 66. A cultured cell into which has been introduced an expression vector of claim 62, wherein the cell-expresses the protein encoded by the DNA segment.
- 67. A cultured cell into which has been introduced an expression vector of claim 64, wherein the cell expresses the protein encoded by the DNA segment.
- 68. A method of producing a protein comprising:
culturing a cell into which has been introduced an expression vector of claim 62, whereby the cell expresses the protein encoded by the DNA segment; and recovering the expressed protein.
- 69. A method of producing a protein comprising:
culturing a cell into which has been introduced an expression vector of claim 64, whereby the cell expresses the protein encoded by the DNA segment; and recovering the expressed protein.
- 70. An isolated polynucleotide molecule encoding a polypeptide comprising amino acid residues 23 to 67 of SEQ ID NO: 10.
REFERENCE TO RELATED APPLICATIONS
[0001] This application is continuation of U.S. patent application Ser. No. 09/636,399, filed on Aug. 10, 2000, which is a continuation-in-part of U.S. patent application Ser. No. 09/344,097, filed on Jun. 25, 1999, which is a continuation-in-part of U.S. patent application Ser. No. 09/150,786, filed on Sep. 10, 1998, which claims the benefit of U.S. Provisional Application Serial No. 60/058,335, filed on Sep. 10, 1997 and U.S. Provisional Application Serial No. 60/064,294, filed on Nov. 5, 1997, all of which are herein incorporated by reference.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60058335 |
Sep 1997 |
US |
|
60064294 |
Nov 1997 |
US |
Continuations (1)
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Number |
Date |
Country |
Parent |
09636399 |
Aug 2000 |
US |
Child |
10409532 |
Apr 2003 |
US |
Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
09344097 |
Jun 1999 |
US |
Child |
09636399 |
Aug 2000 |
US |
Parent |
09150786 |
Sep 1998 |
US |
Child |
09344097 |
Jun 1999 |
US |