1. In a reproducing apparatus comprising a loading unit arranged to move a medium holding member from a closed position where a reproducing operation can be performed to an opened position where a medium is to be mounted and removed,
the reproducing apparatus comprising:
an open key for inputting an open command for causing the loading unit to move the medium holding member from the closed position to the opened position;
a setting unit arranged to allow a user to set the open key press duration, required for the loading unit to perform an opening operation, to the shortest time period or a longer time period than the shortest time period; and
a control unit arranged to cause, in a case where the open key is operated, the loading unit to perform the opening operation when the open key press duration reaches the open key press duration set by the setting unit,
wherein the user is a human operator,
said reproducing apparatus further comprising:
a unit arranged to cause, in a case where the setting unit sets the longer time period than the shortest time period as the open key press duration, a display device to indicate that the opening operation performed by the loading unit is restricted only when the open key is pressed down.
2. The reproducing apparatus according to claim 1, wherein the shortest time period is 3 seconds and the longest time period is 10 seconds.
3. The reproducing apparatus according to claim 1, wherein the open key press duration set by the setting unit is selected on a menu screen displayed on the display device.
4. The reproducing apparatus according to claim 3, wherein the reproducing apparatus is a Digital Versatile Disc (DVD) player, a Mini Disc (MD) player, a Compact Disc (CD) player, or Video Tape Recorder (VTR).
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. Nucleic acid sequence coding for a thymidine kinase, characterized in that it possesses, relative to the wild-type sequence, at least one mutation in the region corresponding to the ATP-binding site combined with at least one mutation in the N-terminal andor C-terminal region.
2. Nucleic acid sequence, characterized in that it is derived from the sequence coding for a wild-type thymidine kinase, the said sequence possessing at least one mutation in the region corresponding to the ATP-binding site and at least one mutation in the N-terminal andor C-terminal region.
3. Nucleic acid sequence according to claim 2, characterized in that it is derived from the sequence coding for herpes simplex virus type 1 thymidine kinase.
4. Nucleic acid sequence according to claim 3, characterized in that the said sequence comprises at least one substitution of a guanine at position 180 by an adenine (G180A).
5. Nucleic acid sequence according to claim 3 or 4, characterized in that the said sequence comprises at least one substitution of a guanine at position 180 by an adenine (G180A) and at least one substitution of the guanine at position 16 by an adenine (G16A).
6. Nucleic acid sequence according to claim 3 or 4, characterized in that the said sequence comprises at least one substitution of a guanine at position 180 by an adenine (G180A) and at least a double substitution of the guanines at position 28 and 30 by adenines (G28A and G30A).
7. Nucleic acid sequences according to one of claims 4 to 6, characterized in that the said sequence comprises, in addition, at least one mutation in its C-terminal region.
8. Nucleic acid sequence according to one of claims 4, 6 and 7, characterized in that it possesses at least one substitution of a guanine at position 180 by an adenine (G180A), at least a double substitution of the guanines at position 28 and 30 by adenines (G28A and G30A) a double substitution of the cystosines at position 591 and 892 by thymines (C591T and C892T) and a double substitution of the guanines at position 1010 and 1011 by adenines (G1010A and G1011A).
9. Nucleic acid sequence coding for a thymidine kinase variant, characterized in that it is chosen from:
(a) the sequence SEQ ID No. 3 or a portion of the latter carrying the (G180A) mutation or one of their complementary strand,
(b) the sequences SEQ ID No. 6 and SEQ ID No. 7 or a portion of these sequences carrying the (G180A) mutation and, respectively, the G16A mutation and the (G28A; G30A) double mutation or one of their complementary strand,
(c) the sequence SEQ ID No. 8 or a portion of the latter carrying the (G180A) mutation the (G28A: G30A) double mutation, and the (C591T; C892T; G1010A; G1011A) quadruple mutation or one of their complementary strand
(d) any sequence hybridizing with the sequences (a), (b) andor (c) and coding for a thymidine kinase variant and as defined in claim 1 or 2,
(e) the variants of (a), (b), (c) and (d) resulting from the degeneracy of the genetic code.
10. Nucleic acid sequence coding for a variant of a wild-type thymidine kinase, capable of being obtained by mutagenesis, site-directed or otherwise, of a sequence according to one of claims 1 to 9.
11. Nucleic acid sequence according to one of the preceding claims, characterized in that it can be of eukaryotic, bacterial, viral, synthetic or semi-synthetic origin.
12. Variant of a wild-type thymidine kinase, capable of being expressed from a nucleic acid sequence according to any one of claims 1 to 11.
13. Variant of a thymidine kinase, comprising at least one mutation in the region corresponding to the ATP-binding site combined with at least one mutation in its N-terminal andor C-terminal region.
14. Variant according to claim 13, characterized in that the region involved in the region corresponding to the ATP-binding site is represented by the consensus GXXXXGK(TS).
15. Variant according to claim 14, characterized in that it is preferably the motif GPHGMGKT.
16. Variant according to claim 15, characterized in that it comprises at least one substitution at position 60 of a methionine by an isoleucine.
17. Variant of a wild-type thymidine kinase, characterized in that it is the mutant 1537:E4.
18. Variant according to one of claims 13 to 16, characterized in that the mutation in the N-terminal region lies between amino acids 1 and 20 of the said region.
19. Variant according to claim 18, characterized in that this mutation lies between amino acids 1 and 15 of the N-terminal region.
20. Variant according to claim 19, characterized in that this mutation lies between amino acids 1 and 10 of the N-terminal region.
21. Variant according to claim 20, characterized in that this mutation lies between amino acids 5 and 10 of the N-terminal region.
22. Variant of herpes simplex virus type 1 thymidine kinase comprising at least one substitution at position 60 of a methionine by an isoleucine and a substitution at position 10 of an alanine by a threonine.
23. Variant of herpes simplex virus type 1 thymidine kinase comprising at least one substitution at position 60 of a methionine by an isoleucine and a substitution at position 6 of a glycine by a serine.
24. Variant of a wild-type thymidine kinase, characterized in that it is the mutant 2-865:H12.
25. Variant of a wild-type thymidine kinase, characterized in that it is the mutant 2-3361:D3.
26. Variant according to claims 14 to 23, comprising, in addition, at least one mutation in the C-terminal region.
27. Variant according to claim 26, characterized in that this mutation lies between amino acids 320 and 350 of the C-terminal region.
28. Variant according to claim 27, characterized in that this mutation lies between amino acids 325 and 345 of the C-terminal region.
29. Variant according to claim 28, characterized in that this mutation lies between amino acids 330 and 343 of the C-terminal region.
30. Variant according to claim 29, characterized in that this mutation lies between amino acids 335 and 340 of the C-terminal region.
31. Variant of herpes simplex virus type 1 thymidine kinase comprising at least one substitution at position 60 of a methionine by an isoleucine, a substitution at position 10 of an alanine by a threonine and a substitution at position 337 of an arginine by a glutamine.
32. Variant of a wild-type thymidine kinase, characterized in that it is the mutant 3-4216:H2.
33. Variant of a wild-type thymidine kinase, characterized in that it displays the following kinetic properties:
a substantial or even complete decrease in the inhibition of the activity of phosphorylation of ganciclovir contrary to the wild-type enzyme for which the inhibition is very marked at and above 15 M;
an increase by at least a factor of 2 to 2.5 in the initial rate of phosphorylation of GCV at and above 15-20 M, relative to the wild-type enzyme and
a KcatKm ratio for thymidine which is reduced by a factor of 1 to 6 relative to that of the wild-type enzyme.
34. Variant of a thymidine kinase according to claims 13 to 25, characterized in that it displays at least one of the following kinetic performance features:
a significant decrease in the inhibition of the phosphorylation of ganciclovir or of the nucleoside analogue at high concentrations of ganciclovir or of nucleoside analogue;
a rate of phosphorylation of ganciclovir or of nucleoside analogue, which is at least tripled, andor
a KcatKm ratio comes out unchanged either for thymidine reduced by a factor greater than or equal to 5 relative to that of the wild-type enzyme.
35. Variant of a thymidine kinase according to claims 26 to 32, characterized in that it displays at least one of the following kinetic performance features:
an absence of inhibition of the phosphorylation of ganciclovir or of the nucleoside analogue at high concentrations of ganciclovir or of nucleoside analogue,
an increase by a factor greater than 3.5 in the initial rate of phosphorylation of GCV at and above 15-20 M, relative to the wild-type enzyme.
a KcatKm ratio for thymidine which is decreased by a factor of 4, relative to that of the wild-type enzyme.
36. Expression cassette comprising a nucleic acid according to one of claims 1 to 11, a promoter permitting its expression and a transcription termination signal.
37. Vector comprising a nucleic acid according to one of claims 1 to 11 or a cassette according to claim 36.
38. Vector according to according to claim 37, characterized in that it is a viral vector.
39. Vector according to claim 38, characterized in that it is a defective recombinant adenovirus.
40. Vector according to claim 38, characterized in that it is a defective recombinant retrovirus.
41. Vector according to claim 38, characterized in that it is a defective recombinant AAV.
42. Vector according to claim 38, characterized in that it is a defective recombinant HSV.
43. Vector according to claim 37, characterized in that it is a chemical or biochemical vector.
44. Pharmaceutical composition comprising a nucleic acid according to any one of claims 1 to 11, andor a vector according to one of claims 37 to 43.
45. Pharmaceutical composition comprising a variant according to one of claims 12 to 35.
46. Pharmaceutical composition according to either of claims 45 and 46, for the treatment of hyperproliferative disorders.