Claims
- 1. A method of effecting a change in a cell, wherein a transfer system is contacted with the cell to be changed, said transfer system comprising a membrane with in it a protein transport system comprising a pore which comprises a VirB complex and VirD4 protein, wherein the transfer system comprises a fusion protein or is capable of making a fusion protein which is introduced into the cell using the protein transport system,
wherein a fusion protein BA is introduced into the cell to be changed which fusion protein BA comprises
i) as a first part A, an oligopeptide comprising the C-terminal amino acids 1-20 of VirF, VirD2, VirE2, VirE3, VirD5 or MobA, or an analogue thereof and ii) as a second part B, a polypeptide capable of exerting a cell-changing activity in the cell to be changed, wherein the C-terminal end of the polypeptide is linked to the N-terminal end of the first part A, under the condition that if the fusion protein comprises a first part A derived from VirE2, the fusion protein does not comprise the 84 N-terminal amino acids of VirE2.
- 2. The method according to claim 1, wherein the oligopeptide of the part A comprises at least two residues from the group consisting essentially of arginine and lysine, and that it contains 50% or more hydrophilic or neutral amino acids.
- 3. The method according to claim 1, wherein the fusion protein is formed by expression in the transfer system.
- 4. The method according to claim 2, wherein the fusion protein is formed by expression in the transfer system.
- 5. The method according to claim 1, wherein the fusion protein is introduced into the cell to be changed without introducing a DNA- or RNA-sequence.
- 6. The method according to claim 2, wherein the fusion protein is introduced into the cell to be changed without introducing a DNA- or RNA-sequence.
- 7. The method according to claim 3, wherein the fusion protein is introduced into the cell to be changed without introducing a DNA- or RNA-sequence.
- 8. The method according to claim 4, wherein the fusion protein is introduced into the cell to be changed without introducing a DNA- or RNA-sequence.
- 9. The method according to claim 1, wherein the introduced fusion protein possesses a recombinase activity.
- 10. The method according to claim 2, wherein the introduced fusion protein possesses a recombinase activity.
- 11. The method according to claim 3, wherein the introduced fusion protein possesses a recombinase activity.
- 12. The method according to claim 4, wherein the introduced fusion protein possesses a recombinase activity.
- 13. The method according to claim 5, wherein the introduced fusion protein possesses a recombinase activity.
- 14. The method according to claim 6, wherein the introduced fusion protein possesses a recombinase activity.
- 15. The method according to claim 7, wherein the introduced fusion protein possesses a recombinase activity.
- 16. The method according to claim 8, wherein the introduced fusion protein possesses a recombinase activity.
- 17. The method according to any one of claims 1 through 17, wherein a bacterium from the class of the Rhizobiaceae is used as the transfer system.
- 18. The method according to any one claims 1 through 18, wherein the cell to be modified is selected from the group consisting of i) a plant cell; ii) a yeast cell; and iii) a fungal cell.
- 19. A vector that it codes for a protein transport system comprising a pore which contains a VirB complex and VirD4 protein, as well as for a fusion protein BA that comprises i) as a first part A an oligopeptide comprising the C-terminal amino acids 1-20 of VirF, VirD2, VirE2, VirE3, VirD5 or MobA, or an analogue thereof ii) as a second part B a polypeptide capable of effecting a cell-modifying activity in the cell to be modified, wherein the polypeptide with its C-terminal end is linked to the N-terminal end of the first part A, under the condition that if the fusion protein comprises a first part A derived from VirE2, the fusion protein does not comprise the 84 N-terminal amino acids of VirE2.
- 20. A vector set, characterized in that the vector set comprises one or more vectors coding for a protein transport system comprising a pore which contains a VirB complex and VirD4 protein as well as a further vector coding for a fusion peptide BA which comprises i) as a first part A an oligopeptide comprising the C-terminal amino acids 1-20 of VirF, VirD2, VirE2, VirE3, VirD5 or MobA, or an analogue thereof, and ii) as a second part B a polypeptide capable of exercising a cell-modifying activity in the cell to be modified, wherein the polypeptide of the C-terminal end of the polypeptide is linked to the N-terminal end of the first part A, under the condition that if the fusion protein comprises a first part A derived from VirE2, the fusion protein does not comprise the 84 N-terminal amino acids of VirE2.
Priority Claims (2)
Number |
Date |
Country |
Kind |
WO 01/89283 A1 |
Nov 2001 |
WO |
|
1015252 |
May 2000 |
NL |
|
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part application of PCT International Application No. PCT/NL01/00388, filed on May 21, 2001 and designating the United States of America, published in English on Nov. 21, 2001 as WO 01/89283 A1, the contents of the entirety of which are incorporated by this reference.
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
PCT/NL01/00388 |
May 2001 |
US |
Child |
10300666 |
Nov 2002 |
US |