Claims
- 1. An expression vector comprising a nucleic acid encoding for at least one of the proteins selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8.
- 2. A host cell comprising the expression vector of claim 1.
- 3. An expression vector comprising a nucleic acid encoding for at least one of the genes selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13.
- 4. A host cell comprising the expression vector of claim 3.
- 5. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 1 or a degenerate variant of SEQ ID NO: 1.
- 6. An isolated nucleic acid molecule comprising a sequence that encodes a polypeptide having amino acid sequence SEQ ID NO: 2, or SEQ ID NO: 2 with conservative amino acid substitutions.
- 7. A method of preparing a protein having amino acid sequence SEQ ID NO:2, or SEQ ID NO: 2 with conservative amino acid substitutions comprising culturing the host cell of claim 2 under conditions permitting the expression of said protein and recovering said protein from the cell culture.
- 8. A purified polypeptide molecule comprising the amino acid sequence SEQ ID NO: 2 or SEQ ID NO: 2 with conservative amino acid substitutions.
- 9. An antibody that binds specifically to Aap.
- 10. The antibody of claim 9, wherein said antibody is selected from the group consisting of polyclonal, monoclonal, chimeric, single chain and Fab fragments.
- 11. The antibody of claim 9, wherein said antibody is detectably labeled.
- 12. The antibody of claim 11, wherein said label is selected from the group consisting of a radioisotope, a bioluminescent compound, a chemiluminescent compound, a fluorescent compound, a metal chelate and an enzyme.
- 13. A method of detecting the presence of Aap in a sample comprising the steps of:
contacting said sample with the antibody of claim 9; and detecting the binding of said antibody to Aap; wherein said binding is indicative of the presence of Aap in said sample.
- 14. The method of claim 13, wherein said presence of Aap is indicative of the presence of enteroaggregative E. coli.
- 15. The method of claim 13, wherein said sample is hair, skin, tissue, cultured cells, cultured cell media and biological fluids.
- 16. The method of claim 13, wherein said antibody is labeled.
- 17. The method of claim 16, wherein said label is selected from the group consisting of a radioisotope, a bioluminescent compound, a chemiluminescent compound, a fluorescent compound, a metal chelate and an enzyme.
- 18. A kit comprising a vessel containing the antibody of claim 9 and instructions for using the antibody to detect protein Aap.
- 19. A kit for detecting an infection caused by enteroaggregative E. coli comprising:
a container, a composition containing the antibody of claim 9 within said container, a visualization means for detecting binding of the antibody to Aap; and instructions for using the composition to detect an infection caused by enteroaggregative E. coli; wherein the label on said container indicates that the composition can be used for the detection of an infection caused by enteroaggregative E. coli.
- 20. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 9 or a degenerate variant of SEQ ID NO: 9.
- 21. An isolated nucleic acid molecule comprising a sequence that encodes a polypeptide having amino acid sequence SEQ ID NO: 4, or SEQ ID NO: 4 with conservative amino acid substitutions.
- 22. A method of preparing a protein having amino acid sequence SEQ ID NO:4, or SEQ ID NO: 4 with conservative amino acid substitutions comprising culturing the host cell of claim 2 under conditions permitting the expression of said protein and recovering said protein from the cell culture.
- 23. A purified polypeptide molecule comprising the amino acid sequence SEQ ID NO: 4 or SEQ ID NO: 4 with conservative amino acid substitutions.
- 24. An antibody that binds specifically to AatP.
- 25. A method of detecting the presence of protein AatP in a sample comprising the steps of:
contacting said sample with the antibody of claim 24; and detecting the binding of said antibody to AatP; wherein said binding is indicative of the presence of AatP in said sample.
- 26. The method of claim 25, wherein said presence of AatP is indicative of the presence of enteroaggregative E. coli.
- 27. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 10 or a degenerate variant of SEQ ID NO: 10.
- 28. An isolated nucleic acid molecule comprising a sequence that encodes a polypeptide having amino acid sequence SEQ ID NO: 5, or SEQ ID NO: 5 with conservative amino acid substitutions.
- 29. A method of preparing a protein having amino acid sequence SEQ ID NO:5, or SEQ ID NO: 5 with conservative amino acid substitutions comprising culturing the host cell of claim 2 under conditions permitting the expression of said protein and recovering said protein from the cell culture.
- 30. A purified polypeptide molecule comprising the amino acid sequence SEQ ID NO: 5 or SEQ ID NO: 5 with conservative amino acid substitutions.
- 31. An antibody that binds specifically to AatA.
- 32. A method of detecting the presence of AatA in a sample comprising the steps of:
contacting said sample with the antibody of claim 31; and detecting the binding of said antibody to AatA; wherein said binding is indicative of the presence of AatA in said sample.
- 33. The method of claim 32, wherein said presence of AatA is indicative of the presence of enteroaggregative E. coli.
- 34. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 11 or a degenerate variant of SEQ ID NO: 11.
- 35. An isolated nucleic acid molecule comprising a sequence that encodes a polypeptide having amino acid sequence SEQ ID NO: 6, or SEQ ID NO: 6 with conservative amino acid substitutions.
- 36. A method of preparing a protein having amino acid sequence SEQ ID NO:6, or SEQ ID NO: 6 with conservative amino acid substitutions comprising culturing the host cell of claim 2 under conditions permitting the expression of said protein and recovering said protein from the cell culture.
- 37. A purified polypeptide molecule comprising the amino acid sequence SEQ ID NO: 6 or SEQ ID NO: 6 with conservative amino acid substitutions.
- 38. An antibody that binds specifically to AatB.
- 39. A method of detecting the presence of protein AatB in a sample comprising the steps of:
contacting said sample with the antibody of claim 38; and detecting the binding of said antibody to AatB; wherein said binding is indicative of the presence of AatB in said sample.
- 40. The method of claim 39, wherein said presence of AatB is indicative of the presence of enteroaggregative E. coli.
- 41. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 12 or a degenerate variant of SEQ ID NO: 12.
- 42. An isolated nucleic acid molecule comprising a sequence that encodes a polypeptide having amino acid sequence SEQ ID NO: 7, or SEQ ID NO: 7 with conservative amino acid substitutions.
- 43. A method of preparing a protein having amino acid sequence SEQ ID NO:7, or SEQ ID NO:7 with conservative amino acid substitutions comprising culturing the host cell of claim 2 under conditions permitting the expression of said protein and recovering said protein from the cell culture.
- 44. A purified polypeptide molecule comprising the amino acid sequence SEQ ID NO: 7 or SEQ ID NO: 7 with conservative amino acid substitutions.
- 45. An antibody that binds specifically to AatC.
- 46. A method of detecting the presence of AatC in a sample comprising the steps of:
contacting said sample with the antibody of claim 45; and detecting the binding of said antibody to AatC; wherein said binding is indicative of the presence of AatC in said sample.
- 47. The method of claim 46, wherein said presence of AatC is indicative of the presence of enteroaggregative E. coli.
- 48. An isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 13 or a degenerate variant of SEQ ID NO: 13.
- 49. An isolated nucleic acid molecule comprising a sequence that encodes a polypeptide having amino acid sequence SEQ ID NO: 8, or SEQ ID NO: 8 with conservative amino acid substitutions.
- 50. A method of preparing a protein having amino acid sequence SEQ ID NO: 8, or SEQ ID NO: 8 with conservative amino acid substitutions comprising culturing the host cell of claim 2 under conditions permitting the expression of said protein and recovering said protein from the cell culture.
- 51. A purified polypeptide molecule comprising the amino acid sequence SEQ ID NO: 8 or SEQ ID NO: 8 with conservative amino acid substitutions.
- 52. An antibody that binds specifically to AatD.
- 53. A method of detecting the presence of AatD in a sample comprising the steps of:
contacting said sample with the antibody of claim 52; and detecting the binding of said antibody to AatD; wherein said binding is indicative of the presence of AatD in said sample.
- 54. The method of claim 53, wherein said presence of AatD is indicative of the presence of enteroaggregative E. coli.
- 55. An immunogenic composition capable of stimulating an immune response in an animal to enteroaggregative E. coli comprising a pharmaceutically acceptable carrier and one or more of the isolated, purified proteins selected from the group consisting of Aat, AatP, AatA, AatB, AatC, and AatD or fragments thereof.
- 56. The immunogenic composition of claim 55, wherein said immunogenic composition is administered by a route selected from the group consisting of oral administration, nasal administration, inhalation, ophthalmic administration, rectal administration and vaginal administration.
- 57. A method of generating an immune response to enteroaggregative E. coli in an animal comprising administering the immunogenic composition of claim 55 to the animal.
- 58. An immunogenic composition capable of stimulating an immune response in an animal to enteroaggregative E. coli comprising a pharmaceutically acceptable carrier and one or more isolated nucleic acid molecules which encode a polypeptide selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, or these sequences with conservative amino acid substitutions.
- 59. The immunogenic composition of claim 58, wherein said immunogenic composition is administered by a route selected from the group consisting of oral administration, nasal administration, inhalation, ophthalmic administration, rectal administration and vaginal administration.
- 60. A method of generating an immune response to enteroaggregative E. coli in an animal comprising administering the immunogenic composition of claim 58 to the animal.
- 61. A kit comprising:
a container, the immunogenic composition of claim 55 contained within said container, and instructions for using the immunogenic composition to elicit an immune response to enteroaggregative E. coli; wherein said label on said container indicates that said immunogenic composition can be used to elicit an immune response in an animal to enteroaggregative E. coli.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This non-provisional application claims priority under 35 U.S.C. §120 to provisional applications U.S. Serial Nos. 60/334,425 and 60/398,775, filed Nov. 30, 2001 and Jul. 26, 2002, respectively, the contents of which are incorporated herein by reference in their entirety.
STATEMENT OF GOVERNMENTAL INTEREST
[0002] This invention was made with U.S. Government support under NIADA grant No. AI133096 awarded by the National Institutes of Health. The U.S. Government may have certain rights in this invention.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60334425 |
Nov 2001 |
US |
|
60398775 |
Jul 2002 |
US |