Claims
- 1. A human factor IX molecule of formula (Ala).sub.1 FIX wherein FIX refers to natural or recombinant human factor IX.
- 2. A DNA fragment which encodes for the human factor IX polypeptide of claim 1.
- 3. A DNA fragment which comprises at least:
- a first portion encoding the human factor IX molecule of claim 1, and
- a second portion linked to the 5' end of said first portion and encoding a prosequence, which prosequence provides for carboxylation of the human factor IX molecule and is removed by cleavage upon secretion of the human factor IX molecule, which has at position -3 a valine, arginine, lysine, threonine or serine residue.
- 4. The DNA fragment of claim 3 wherein said prosequence is derived from the prosequence of the human factor IX precursor and differs therefrom in that the native proline at position -3 is replaced by a valine, arginine, lysine, threonine or serine residue.
- 5. The DNA fragment of claim 3 further comprising a third portion linked to the 5' end of said second portion and encoding a signal peptide.
- 6. The DNA fragment of claim 5 wherein the signal peptide is the signal peptide of the human factor IX precursor.
- 7. The DNA fragment of claim 5 wherein the signal peptide is a vitamin K-dependent protein signal peptide.
- 8. A plasmid which contains the DNA fragment of claim 2 under control of elements which provide for the expression of said DNA fragment.
- 9. A poxvirus vector which contains the DNA fragment of claim 2 under the control of elements providing for the expression of said DNA fragment.
- 10. The poxvirus vector of claim 9 which is a vaccinia virus.
- 11. A cell transformed by the plasmid of claim 8.
- 12. A cell culture infected with the viral vector of claim 9.
- 13. A cell which has inserted in its genome the DNA fragment of claim 2 under the control of elements providing for the expression of said DNA fragment.
- 14. The cell of claim 11 or 13 wherein the cell is a mammalian cell.
- 15. The cell culture of claim 9 wherein the cell culture is a mammalian cell culture.
- 16. A method for preparing (Ala).sub.1 FIX comprising culturing the cell or cell culture of any one of claims 11, 12 or 13, and recovering (Ala).sub.1 FIX from the culture.
Priority Claims (1)
Number |
Date |
Country |
Kind |
88 14635 |
Nov 1988 |
FRX |
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Parent Case Info
This application is a continuation of application Ser. No. 07/970,966, filed Nov. 3, 1992, now abandoned, which is a file wrapper continuation of Ser. No. 07/433,276, filed Nov. 8, 1989, now abandoned.
US Referenced Citations (2)
Number |
Name |
Date |
Kind |
4760025 |
Estell |
Jul 1988 |
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4770999 |
Kaufman |
Sep 1988 |
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Non-Patent Literature Citations (5)
Entry |
Pavirani et al. Biotechnology 5:384, 1987. |
Kurochi et al PNAS 79: 6461, 1982. |
Bentley et al Cell 45: 343, 1986. |
DiScipio et al Biochemistry 16(4): 698, 1977. |
de la Salle et al Nature 316: 268, 1985. |
Continuations (2)
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Number |
Date |
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Parent |
970966 |
Nov 1992 |
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Parent |
433276 |
Nov 1989 |
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