Claims
- 1. A recombinant DNA sequence encoding an analog of mammalian FGF.
- 2. The DNA sequence of claim 1 which encodes a human FGF protein analog.
- 3. The DNA sequence of claim 2 which encodes a human basic FGF protein analog.
- 4. The DNA sequence of claim 3 which encodes a human basic FGF protein analog with reduced affinity for heparin binding.
- 5. The DNA sequence of claim 4 encoding a human basic FGF protein analog, comprising substituting one or more positively charged amino acid residues located in a heparin binding domain encompassing residues 128 through 138 with a neutral or negatively charged amino acid.
- 6. The DNA sequence of claim 5 wherein the neutral or negatively charged amino acid is selected from the group consisting of serine, threonine or glutamic acid.
- 7. The DNA sequence of claim 5 wherein the location and composition of the substituted amino acid is selected from the group consisting of serine128, glutamic acid128, threonine129, serine128/threonine129, and serine138.
- 8. The DNA sequence of claim 2 which encodes a human basic FGF protein analog wherein one or more cysteine residues are replaced by a neutral amino acid and said protein analog exhibits the biological activity of native basic FGF.
- 9. The DNA sequence of claim 8 wherein the neutral amino acid is serine or alanine.
- 10. The DNA sequence of claim 9 wherein the substituted cysteine residue is at position 78, 96 or a combination thereof.
- 11. The DNA sequence of claim 5 which encodes a human basic FGF protein analog wherein said analog binds to a receptor for FGF and has reduced ability to induce a biological response.
- 12. The DNA sequence of claim 3 which encodes an amino-terminal deletion analog of FGF having FGF antagonist activity.
- 13. The DNA sequence of claim 12 wherein said deletion spans residues 1 through 24 of human basic FGF.
- 14. The DNA sequence of claim 12 encoding a human basic FGF analog further comprising one or more positively charged amino acid residues located in a heparin binding domain encompassing residues 128 through 138 substituted with a neutral or negatively charged amino acid.
- 15. The DNA sequence of claim 3 which is operably linked to control sequences for expression.
- 16. The DNA sequence of claim 15 wherein the control sequences include a transcription termination signal.
- 17. The DNA sequence of claim 3 which is transformed into a recombinant host cell.
- 18. A recombinant vector containing the DNA sequence of claim 3 and effective in expressing FGF or an analog thereof.
- 19. The vector of claim 18 which is selected from the group consisting of plasmids pUC9-TSF11 and pUC9delH3-pTSF-3.
- 20. The vector of claim 18 wherein the DNA sequence encoding an FGF analog is operably linked to control sequences compatible with bacteria.
- 21. The vector of claim 18 wherein the DNA sequence encoding an FGF analog is operably linked to control sequences compatible with mammalian hosts.
- 22. Recombinant host cells transformed with the vector of claim 18.
- 23. Bacterial cells transformed with the vector of claim 20.
- 24. Mammalian cells transformed with the vector of claim 21.
- 25. A method for producing FGF protein analogs which comprises culturing host cells harboring the DNA of claim 3 and recovering the FGF protein analog.
- 26. The method of claim 25 wherein the host cells are bacterial.
- 27. The method of claim 25 wherein the host cells are mammalian.
- 28. A human basic FGF protein analog having reduced affinity for heparin binding comprising substituting one or more positively charged amino acid residues located in a heparin binding domain encompassing residues 128 through 138 with a neutral or negatively charged amino acid.
- 29. A human basic FGF protein analog wherein the cysteine at positions 78, 96, or a combination thereof, is replaced by a neutral amino acid and said analog exhibits the biological activity of native, human basic FGF.
- 30. The human basic FGF protein analog of claim 29 which is bFGF-C78/96S.
- 31. An antagonist of human basic FGF.
- 32. The FGF antagonist of claim 31 wherein the first 24 amino terminal residues of basic FGF are deleted.
Priority Claims (1)
Number |
Date |
Country |
Kind |
518137 |
Dec 1986 |
CA |
|
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This is a continuation-in-part of U.S. Ser. No. 070,797, filed Jul. 7, 1987, which is a continuation-in-part of U.S. Ser. No. 050,706, filed May 15, 1987, which is a continuation-in-part of U.S. Ser. No. 869,382 filed May 30, 1986, which is a continuation-in-part of U.S. Ser. No. 809,163, filed Dec. 16, 1985, which is a continuation-in-part of U.S. Ser. No. 775,521, filed Sep. 12, 1985.
Divisions (2)
|
Number |
Date |
Country |
Parent |
09098628 |
Jun 1998 |
US |
Child |
09902460 |
Jul 2001 |
US |
Parent |
07459739 |
Feb 1990 |
US |
Child |
09098628 |
Jun 1998 |
US |
Continuation in Parts (5)
|
Number |
Date |
Country |
Parent |
07070797 |
Jul 1987 |
US |
Child |
07459739 |
Feb 1990 |
US |
Parent |
07050706 |
May 1987 |
US |
Child |
07070797 |
Jul 1987 |
US |
Parent |
06869382 |
May 1986 |
US |
Child |
07050706 |
May 1987 |
US |
Parent |
06809163 |
Dec 1985 |
US |
Child |
06869382 |
May 1986 |
US |
Parent |
06775521 |
Sep 1985 |
US |
Child |
06809163 |
Dec 1985 |
US |