US6153745A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Relating to mutagenesis of nucleic acids
Assignee: Medical Research Council
•
Filing: 1996-09-19
•
Granted: 2000-11-28
•
Expires: 2016-09-19
Abstract
Abstract PCT No. PCT/GB96/02333 Sec. 371 Date Jul. 6, 1998 Sec. 102(e) Date Jul. 6, 1998 PCT Filed Sep. 19, 1996 PCT Pub. No. WO97/11083 PCT Pub. Date Mar. 27, 1997The invention concerns novel compounds having defined structural formulae and methods of mutating a nucleic acid sequence, the method comprising replicating a template sequence in the presence of a nucleoside triphosphate analogue in accordance with the invention, so as to form non-identical copies of the template sequence comprising one or more nucleoside triphosphate analogue residues, and a kit for use in performing the method of the invention.
Inventors
- 👤 David Williams
- 👤 Daniel Brown
- 👤 Manuella Carla Zaccolo
- 👤 Ermanno Gherardi
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 23:45
CPC Classifications
C; C07; C07H; C07H19/00; C07H19/02; C07H19/04; C07H19/06; C07H19/10; C07H19/16; C07H19/20; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/102
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
82 words
A compound having the structure: ##STR4## where X 1 is selected from the group consisting of O, S, N-alkyl, N + -dialkyl and N-benzyl; X 2 is selected from the group consisting of triphosphate (P 3 O 9 ) 4- , diphosphate (P 2 O 6 ) 3- and thiotriphosphate (P 3 O 8 S) 4- ; and X 3 is selected from the group consisting of H, NH 2 , F and OR, where R is H, methyl, allyl or alkaryl.
Claim 2
Dependent Claim
82 words
A compound having the structure: ##STR4## where X 1 is selected from the group consisting of O, S, N-alkyl, N + -dialkyl and N-benzyl; X 2 is selected from the group consisting of triphosphate (P 3 O 9 ) 4- , diphosphate (P 2 O 6 ) 3- and thiotriphosphate (P 3 O 8 S) 4- ; and X 3 is selected from the group consisting of H, NH 2 , F and OR, where R is H, methyl, allyl or alkaryl.
Claim 3
Dependent Claim
11 words
A compound according to claim 1, wherein X 1 is O.
Claim 4
Dependent Claim
18 words
A compound according to claim 2, wherein, X 2 is triphosphate, and X 3 is H or OH.
Claim 5
Dependent Claim
44 words
A method of mutating a nucleic acid sequence, comprising replicating a template sequence in the presence of a nucleoside triphosphate analogue in accordance with claim 1, so as to form non-identical copies of the template sequence having one or more nucleoside phosphate analogue residues.
Claim 6
Dependent Claim
34 words
A method according to claim 4, comprising replicating a template sequence in the presence of deoxyP triphosphate, so as to form non-identical copies of the template sequence having one or more dP nucleotide residues.
Claim 7
Dependent Claim
24 words
A method according to claim 4, further comprising wherein the template sequence is replicated in the presence of one or more additional nucleoside triphosphates.
Claim 8
Dependent Claim
115 words
A method according to claim 4, further comprising wherein the template sequence is replicated in the presence of a second compound having the structure: ##STR5## where Y 1 is selected from the group consisting of OH, O-alkyl, NH 2 and N(Alkyl) 2 ; Y 2 is selected from the group consisting of H and NH 2 ; Y 3 is selected from the group consisting of triphosphate (P 3 O 9 ) 4- , diphosphate (P 2 O 6 ) 3- and thiotriphosphate (P 3 O 8 S) 4- ; and Y 4 is selected from the group consisting of H, NH 2 , F and OR, where R is H, methyl, allyl or alkaryl.
Claim 9
Dependent Claim
36 words
A method according to claim 4, further comprising wherein the template sequence is replicated in the presence of at least one member of the group consisting of 2'-deoxy-8-hydroxyguanosine 5'-triphosphate, 2-amino-9-(2-deoxy-β-D-erythropentofuranosyl)-6-methoxyaminopurine 5'-triphosphate, and O 2 -ethylthymidine triphosphate.
Claim 10
Dependent Claim
49 words
A method according to claim 4, further comprising the step of replicating the non-identical copies of the template sequence in the presence of the four normal dNTPs, but in the absence of analogues thereof, to form further non-identical copies of the template sequence comprising only the four normal deoxynucleotides.
Claim 11
Dependent Claim
27 words
A method according to claim 4, wherein the replication of the template sequence, and/or the replication of the non-identical copies thereof, is achieved by means of PCR.
Claim 12
Dependent Claim
23 words
A method according to claim 4, further comprising wherein the template sequence is replicated in the additional presence of the four normal deoxynucleotides.
Claim 13
Dependent Claim
26 words
A method according to claim 5, wherein the template sequence is replicated in the presence of 1 μM to 600 μM 6-(2-deoxy-β-D-erythropentofuranosyl)-3,4 dihydro-8H-pyrimido [4,5-c][1,2]oxazine-7-one 5' triphosphate.
Claim 14
Dependent Claim
23 words
A method according to claim 5, wherein the template sequence is replicated in the presence of 1 μM to 600 μM 2'-deoxy-8-hydroxyguanosine 5'-triphosphate.
Claim 15
Dependent Claim
48 words
A kit for performing the method of claim 4, comprising said nucleoside triphosphate analogue means for replicating a template sequence so as to incorporate the nucleoside monophosphate portion of the nucleoside triphosphate analogue into non-identical copies of the template sequence, and instructions for use according to said method.
Claim 16
Dependent Claim
16 words
A kit according to claim 14, wherein the nucleoside triphosphate analogue is 6-(2-deoxy-β-D-erythropentofuranosyl)-3,4 dihydro-8H-pyrimido [4,5-c][1,2]oxazine-7-one 5'triphosphate.
Claim 17
Dependent Claim
24 words
A kit according to claim 14, further comprising wherein the means for replicating the template sequence comprises means for performing the polymerase chain reaction.
Claim 18
Dependent Claim
12 words
A kit according to claim 14, further comprising the four normal deoxynucleotides.
Claim 19
Dependent Claim
26 words
A kit according to claim 14, further comprising at least one member of the group consisting of 2'-deoxy-8-hydroxyguanosine 5'-triphosphate, 2-amino-9-(2-deoxy-β-D-erythropentofuranosyl)-6-methoxyaminopurine 5'-triphosphate, and O 2 -ethylthymidine triphosphate.
References Cited (Backward Citations)
US5434257A
Cited
WO1995007918A2
Cited
WO1995011911A1
Cited
WO1997011083A1
Cited
GB9605011D0
Cited
WO1998003673A1
Cited
GB9616425D0
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2003-11-28 | 2004-05-29 | 2004-11-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2007-11-28 | 2008-05-29 | 2008-11-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2011-11-28 | 2012-05-29 | 2012-11-28 | $7700.00 | Window Closed / Lapsed |