Claims
- 1. A library of fusion nucleic acids, each fusion nucleic acid comprising:
a) a first nucleic acid, encoding a scaffold protein sequence; and b) a second nucleic acid, encoding a library peptide sequence comprising an alpha helical biasing sequence; wherein said first nucleic acid is fused to said second nucleic acid.
- 2. A library of fusion nucleic acids, each fusion nucleic acid comprising:
a) a first nucleic acid, encoding a scaffold protein sequence; b) a second nucleic acid, encoding a library peptide sequence; and c) a third nucleic acid, encoding a nucleating sequence; wherein said first nucleic acid is fused to said second nucleic acid, and wherein said second nucleic acid is fused to said third nucleic acid.
- 3. A library according to claim 1 wherein said first nucleic acid is fused to the 5′ end of said second nucleic acid.
- 4. A library according to claim 1 wherein said first nucleic acid is fused to the 3′ end of said second nucleic acid.
- 5. A library according to claim 1 wherein said second nucleic acid is inserted into said first nucleic acid.
- 6. A library according to claim 1 wherein said second nucleic acid comprises nucleic acid sequence encoding random amino acids.
- 7. A library according to claim 2 wherein said second nucleic acid comprises nucleic acid sequence derived from cDNA.
- 8. A library according to claim 1 wherein said library peptide sequence replaces at least one residue of said scaffold protein sequence.
- 9. A library according to claim 2 wherein said third nucleic acid sequence is fused to the 5′ end of said second nucleic acid sequence.
- 10. A library according to claim 1 wherein said first nucleic acid encodes a green fluorescent protein (GFP) protein.
- 11. A library according to claim 10, wherein said second nucleic acid is inserted into said first nucleic acid within positions encoding amino acids 130 to 135.
- 12. A library according to claim 10, wherein said second nucleic acid is inserted into said first nucleic acid within positions encoding amino acids 154 to 159.
- 13. A library according to claim 10, wherein said second nucleic acid is inserted into said first nucleic acid within positions encoding amino acids 172 to 175.
- 14. A library according to claim 10, wherein said second nucleic acid is inserted into said first nucleic acid within positions encoding amino acids 188 to 193.
- 15. A library according to claim 10, wherein said second nucleic acid is inserted into said first nucleic acid within positions encoding amino acids 208 to 216.
- 16. A library according to claim 10 wherein said GFP protein is from Aequorea victoria.
- 17. A library according to claim 16 wherein said library peptide sequence replaces the C-terminal amino acids LGMDELYK (SEQ ID NO: 5) of said Aequorea victoria GFP.
- 18. A library according to claim 10 wherein said GFP protein is from Renilla mulleri.
- 19. A library according to claim 18 wherein said library peptide sequence replaces the C-terminal amino acids LGSLHEWV (SEQ ID NO: 6) of said Renilla mulleri GFP.
- 20. A library according to claim 10 wherein said GFP protein is from Ptilosarcus.
- 21. A library according to claim 1 wherein said first nucleic acid encodes a β-lactamase protein.
- 22. A library according to claim 1 wherein said first nucleic acid encodes a DHFR protein.
- 23. A library according to claim 1 wherein said first nucleic acid encodes a β-galactosidase protein.
- 24. A library according to claim 1 wherein said first nucleic acid encodes a luciferase protein.
- 25. A library according to claim 1 or 2 wherein said fusion nucleic acid further comprises a fourth nucleic acid encoding a linker peptide.
- 26. A library according to claim 25 wherein said fourth nucleic acid is positioned between said first nucleic acid and said second nucleic acid.
- 27. A library according to claim 25 wherein said fusion nucleic acid further comprises a fifth nucleic acid encoding a second linker peptide.
- 28. A library of retroviral vectors comprising the library of fusion nucleic acids according to claim 1.
- 29. A library of host cells comprising the library of fusion nucleic acids according to claim 1.
- 30. A library of host cells comprising the library of retroviral vectors according to claim 28.
- 31. A library of host cells according to claim 29 or 30 wherein said host cells are mammalian host cells.
- 32. A library of fusion polypeptides wherein each polypeptide comprises:
a) a scaffold protein; and b) a library peptide comprising an alpha helical biasing sequence.
- 33. A library of fusion polypeptides wherein each polypeptide comprises:
a) a scaffold protein; b) a library peptide; and c) a nucleating sequence.
- 34. A method of screening for bioactive peptides conferring a particular phenotype comprising:
a) providing a library of cells containing a library of fusion nucleic acids according to claim 1 wherein each said fusion nucleic acid encodes a fusion protein; b) providing conditions under which said fusion proteins are expressed; and c) assaying said cells for said phenotype.
- 35. A method according to claim 34 wherein said providing is accomplished by transfecting said cells with a library of retroviral vectors comprising said library of fusion nucleic acids.
Parent Case Info
[0001] This application is a continuation-in-part of U.S. patent application Ser. No. 09/415,765, filed Oct. 8, 1999, which is a continuation-in-part of U.S. patent application Ser. No. 09/169,015, filed Oct. 8, 1998, now U.S. Pat. No. 6,180,343.
Continuation in Parts (2)
|
Number |
Date |
Country |
Parent |
09415765 |
Oct 1999 |
US |
Child |
10177725 |
Jun 2002 |
US |
Parent |
09169015 |
Oct 1998 |
US |
Child |
09415765 |
Oct 1999 |
US |