Claims
- 1. A method for producing Factor VII or a Factor VII-related polypeptide comprising (a) transfecting a mammalian cell with a nucleic acid molecule comprising (i) a sequence encoding Factor VII or a Factor VII-related polypeptide and (ii) at least one S/MAR element; (b) culturing the transfected cell under conditions suitable for expression of the Factor VII or a Factor VI I-related polypeptide; and (c) isolating the expressed polypeptide.
- 2. The method of claim 1, wherein the nucleic acid molecule comprises two S/MAR elements.
- 3. The method of claim 1, wherein the at least one S/MAR element comprises (i) SEQ ID NO:1 or SEQ ID NO:2, (ii) a functional fragment of SEQ ID NO:1 or SEQ ID NO:2, or (iii) a sequence that is at least about 70% homologous to SEQ ID NO:1 or SEQ ID NO:2.
- 4. The method of claim 2, wherein the two S/MAR elements are selected from (i) SEQ ID NO:1 or SEQ ID NO:2, (ii) functional fragments of SEQ ID NO:1 or SEQ ID NO:2, or (iii) sequences that are at least about 70% homologous to SEQ ID NO:1 or SEQ ID NO:2.
- 5. The method of claim 2, wherein the two S/MAR elements are identical.
- 6. The method of claim 5, wherein the identical S/MAR elements comprise SEQ ID NO:1.
- 7. The method of claim 5, wherein the identical S/MAR elements comprise SEQ ID NO:2.
- 8. The method of claim 5, wherein the identical S/MAR elements comprise SEQ ID NO:3.
- 9. The method of claim 5, wherein the identical S/MAR elements comprise SEQ ID NO:4.
- 10. The method of claim 5, wherein the identical S/MAR elements comprise SEQ ID NO:5.
- 11. The method of claim 2, wherein the two S/MAR elements comprise SEQ ID NO:1 and SEQ ID NO:2, respectively.
- 12. The method of claim 2, wherein the two S/MAR elements comprise SEQ ID NO:2 and SEQ ID NO:3, respectively.
- 13. The method of claim 2, wherein the two S/MAR elements comprise SEQ ID NO:2 and SEQ ID NO:4, respectively.
- 14. The method of claim 2, wherein the two S/MAR elements comprise SEQ ID NO:2 and SEQ ID NO:5, respectively.
- 15. The method of claim 1, wherein the at least one S/MAR element is located less than about 10 kb from the Factor VII or Factor VII-related polypeptide-encoding sequence.
- 16. A method for producing a polypeptide or protein comprising (a) transfecting a mammalian cell with a nucleic acid molecule comprising a (I) sequence encoding the polypeptide or protein and (II) at least one S/MAR element comprising (i) SEQ ID NO:1 or SEQ ID NO:2, (ii) a functional fragment of SEQ ID NO:1 or SEQ ID NO:2, or (iii) a sequence that is at least about 70% homologous to SEQ ID NO:1 or SEQ ID NO:2; (b) culturing the transfected cell under conditions suitable for expression of the polypeptide or protein; and (c) isolating the expressed polypeptide or protein.
- 17. The method of claim 16, wherein the nucleic acid molecule comprises two S/MAR elements.
- 18. The method of claim 17, wherein the two S/MAR elements are identical.
- 19. The method of claim 18, wherein the identical S/MAR elements comprise SEQ ID NO:1.
- 20. The method of claim 18, wherein the identical S/MAR elements comprise SEQ ID NO:2.
- 21. The method of claim 18, wherein the identical S/MAR elements comprise SEQ ID NO:3.
- 22. The method of claim 18, wherein the identical S/MAR elements comprise SEQ ID NO:4.
- 23. The method of claim 18, wherein the identical S/MAR elements comprise SEQ ID NO:5.
- 24. The method of claim 17, wherein the two S/MAR elements comprise SEQ ID NO:1 and SEQ ID NO:2, respectively.
- 25. The method of claim 17, wherein the two S/MAR elements comprise SEQ ID NO:2 and SEQ ID NO:3, respectively.
- 26. The method of claim 17, wherein the two S/MAR elements comprise SEQ ID NO:2 and SEQ ID NO:4, respectively.
- 27. The method of claim 17, wherein the two S/MAR elements comprise SEQ ID NO:2 and SEQ ID NO:5, respectively.
- 28. An isolated DNA molecule comprising one or more S/MAR elements that comprise a sequence selected from the group consisting of SEQ ID NOS:1-5.
- 29. The isolated DNA molecule of claim 28, wherein the DNA molecule comprises a sequence encoding a human protein or polypeptide or a functional analogue of a human protein. or polypeptide.
- 30. The isolated DNA molecule of claim 29, wherein the protein or polypeptide-encoding sequence is located less than about 10 kb from the one or more S/MAR elements.
- 31. The isolated DNA molecule of claim 30, wherein the isolated DNA molecule comprises SEQ ID NO:1.
- 32. The isolated DNA molecule of claim 30, wherein the isolated DNA molecule comprises SEQ ID NO:2.
- 33. The isolated DNA molecule of claim 30, wherein the isolated DNA molecule comprises SEQ ID NO:3.
- 34. The isolated DNA molecule of claim 30, wherein the isolated DNA molecule comprises SEQ ID NO:4.
- 35. The isolated DNA molecule of claim 30, wherein the isolated DNA molecule comprises SEQ ID NO:5.
- 36. A vector construct comprising a nucleic acid molecule comprising (a) a sequence encoding Factor VII or a Factor VII-related polypeptide operably linked to one or more expression control elements and (b) one or more S/MAR elements.
- 37. The vector construct of claim 36, wherein the one or more S/MAR elements comprise a sequence selected from SEQ ID NOS:1-5.
- 38. A mammalian cell comprising the vector of claim 37.
- 39. A vector construct comprising a nucleic acid molecule that comprises (a) a sequence encoding a polypeptide or protein operably linked to one or more expression control elements and (b) at least one S/MAR element comprising a sequence selected from SEQ ID NOS:1-5.
- 40. A mammalian cell comprising the vector of claim 39.
- 41. An isolated DNA molecule consisting essentially of one or more sequences selected from SEQ ID NOS:1-5.
Priority Claims (1)
Number |
Date |
Country |
Kind |
PA 2002 01384 |
Sep 2002 |
DK |
|
CROSS REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This application claims priority under 35 U.S.C. 119 from Danish Patent Application No. PA 2002 01384, filed Sep. 20, 2002, and the benefit of U.S. Provisional Patent Application No. 60/416,566, filed Oct. 7, 2002, the contents of which are fully incorporated herein by reference.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60416566 |
Oct 2002 |
US |