US4681840A
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Deoxyribonucleic acid molecules useful as probes for detecting oncogenes incorporated into chromosomal DNA
Assignee: United States Department of Commerce
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Filing: 1984-01-18
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Granted: 1987-07-21
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Expires: 2004-01-18
Abstract
Abstract A single-stranded deoxyribonucleic acid molecule having a length of less than about 25 kb comprises at least three distinct nucleotide sequences which are the sites for incorporation into a chromosome of a deoxyribonucleic acid molecule encoding a deleterious gene. Deoxyribonucleic acid probes have been prepared from such molecules and are useful as hybridization probes for detecting chromosomal deoxyribonucleic acid which has a deoxyribonucleic acid molecule encoding a deleterious gene, i.e. oncogene, incorporated therein. A single-stranded deoxyribonucleic acid molecule derived from human chromosome 22 which is about 5.8 kb in length contains sites for incorporation of a deoxyribonucleic acid molecule encoding the oncogene c-abl derived from human chromosome 9. Deoxyribonucleic acid probes have been prepared from this molecule and used to detect the abnormal Philadelphia chromosome and chronic myelocytic leukemia.
Inventors
- 👤 John R. Stephenson
- 👤 John Groffen
- 👤 Nora Heisterkamp
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 01:13
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6883; C12Q1/6813; C12Q1/6827; C12Q1/6886; C12Q2600/00; C12Q2600/156
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
46 words
A single-stranded deoxyribonucleic acid molecule having a length less than about 25 kb which comprises at least three distinct nucleotide sequences which are present in a breakpoint cluster region and which are sites for incorporation into a chromosome of a deoxyribonucleic acid molecule encoding an oncogene.
Claim 2
Dependent Claim
46 words
A single-stranded deoxyribonucleic acid molecule having a length less than about 25 kb which comprises at least three distinct nucleotide sequences which are present in a breakpoint cluster region and which are sites for incorporation into a chromosome of a deoxyribonucleic acid molecule encoding an oncogene.
Claim 3
Dependent Claim
18 words
A single-stranded deoxyribonucleic acid molecule according to claim 1, which is less than about 15 kb in length.
Claim 4
Dependent Claim
44 words
A single-stranded deoxyribonucleic acid molecule according to claim 1, wherein the sites for incorporation are breakpoint sites to which a deoxyribonucleic acid molecule which encodes the oncogene derived from a second chromosome, or a distal region of the same chromosome, attaches during a translocation.
Claim 5
Dependent Claim
23 words
A double-stranded deoxyribonucleic acid molecule having a length less than about 25 kb which includes the single-stranded deoxyribonucleic acid molecule of claim 1.
Claim 6
Dependent Claim
16 words
A ribonucleic acid molecule which is complementary to the single-stranded deoxyribonucleic acid molecule of claim 1.
Claim 7
Dependent Claim
19 words
A single-stranded deoxyribonucleic acid molecule of claim 1, wherein the single-stranded deoxyribonucleic acid molecule is derived from a chromosome.
Claim 8
Dependent Claim
16 words
A single-stranded deoxyribonucleic acid molecule according to claim 6 wherein the chromosome is a human chromosome.
Claim 9
Dependent Claim
15 words
A fragment of the deoxyribonucleic acid molecule of claim 1 obtained by restriction enzyme cleavage.
Claim 10
Dependent Claim
10 words
A fragment of claim 8 labelled with a detectable marker.
Claim 11
Dependent Claim
81 words
A method of detecting chromosomal deoxyribonucleic acid containing a deoxyribonucleic acid molecule encoding an omogene incorporated therein which comprises treating total chromosomal deoxyribonucleic acid under denaturing conditions to obtain single-stranded deoxyribonucleic acid molecules, contacting the single-stranded deoxyribonucleic acid molecules so obtained with a deoxyribonucleic acid molecule of claim 9 which includes sites for the incorporation of the oncogene under conditions permitting hybridization of complementary single-stranded molecules and separating hybridized molecules so formed to thereby detect chromosomal deoxyribonucleic acid containing an oncogene.
Claim 12
Dependent Claim
125 words
A method of detecting chromosomal deoxyribonucleic acid containing a deoxyribonucleic acid molecule encoding an oncogene incorporated therein which comprises cleaving total chromosomal deoxyribonucleic acid with restriction enzymes under cleaving conditions, separating the fragments so obtained by gel electrophoresis, denaturing the fragments to obtain single-stranded deoxyribonucleic acid molecules, recovering the denatured single-stranded fragments from the gel, immobilizing them on a solid support, contacting the immobilized single-stranded deoxyribonucleic acid fragments with a deoxyribonucleic acid probe of claim 9 which includes sites for the incorporation of the oncogene under conditions permitting hybridization of complementary single-stranded molecules, identifying the hybridized molecules so formed to thereby detect abnormalities in the restriction patterns of the chromosomal deoxyribonucleic acid caused by the incorporation of deoxyribonucleic acid containing the oncogene into the chromosome.
Claim 13
Dependent Claim
12 words
A deoxyribonucleic acid molecule of claim 1 labelled with a detectable marker.
Claim 14
Dependent Claim
12 words
A deoxyribonucleic acid molecule of claim 12, wherein the marker is radioactive.
Claim 15
Dependent Claim
12 words
A deoxyribonucleic acid molecule of claim 12, wherein the marker is fluorescent.
Claim 16
Dependent Claim
81 words
A method of detecting chromosomal deoxyribonucleic acid containing a deoxyribonucleic acid molecule encoding an oncogene incorporated therein which comprises treating total chromosomal deoxyribonucleic acid under denaturing conditions to obtain single-stranded deoxyribonucleic acid molecules, contacting the single-stranded deoxyribonucleic acid molecules so obtained with a deoxyribonucleic acid molecule of claim 12 which includes sites for the incorporation of the oncogene under conditions permitting hybridization of complementary single-stranded molecules and separating hybridized molecules so formed to thereby detect chromosomal deoxyribonucleic acid containing an oncogene.
Claim 17
Dependent Claim
123 words
A method of detecting chromosomal deoxyribonucleic acid containing a deoxyribonucleic acid molecule encoding an oncogene incorporated therein which comprises cleaving total chromosomal deoxyribonucleic acid with restriction enzymes under cleaving conditions, separating the fragments so obtained by gel electrophoresis, denaturing the fragments to obtain single-stranded deoxyribonucleic acid molecules, recovering the denatured single-stranded fragments from the gel, immobilizing the fragments on a solid support, contacting the immobilized single-stranded deoxyribonucleic acid fragments with a deoxyribonucleic acid probe of claim 12 which includes sites for the incorporation of the oncogene under conditions permitting hybridization of complementary single-stranded molecules, identifying the hybridized molecules so formed to thereby detect abnormalities in the restriction patterns of the chromosomal deoxyribonucleic acid caused by the incorporation of the oncogene into the chromosome.
Claim 18
Dependent Claim
11 words
A method as in claim 16, wherein the oncogene is c-abl.
Claim 19
Dependent Claim
44 words
A single-stranded deoxyribonucleic acid molecule according to claim 1, which is about 5.8 kb long, is derived from human chromosome 22 and contains sites for incorporation of a deoxyribonucleic acid molecule encoding the oncogene c-abl derived from human chromosome 9 during the Philadelphia translocation.
Claim 20
Dependent Claim
12 words
A deoxyribonucleic acid molecule of claim 18 labelled with a detectable marker.
Claim 21
Dependent Claim
134 words
A method of diagnosing chronic myelocytic leukemia in a human subject which is associated with an abnormal Philadelphia chromosome containing the translocated oncogene c-abl which comprises cleaving total chromosomal deoxyribonucleic acid with restriction enzymes under cleaving conditions, separating the fragments so obtained by gel electrophoresis, denaturing the fragments to obtain single-stranded deoxyribonucleic acid molecules, recovering the denatured single-stranded fragments from the gel, immobilizing them on a solid support, contacting the immobilized single-stranded deoxyribonucleic acid fragments with a deoxyribonucleic acid probe of claim 19 which includes sites for the incorporation of the chromosome 9 segment containing the oncogene c-abl under conditions permitting hybridization of complementary single-stranded molecules, identifying the hybridized molecules so formed to thereby detect abnormalities in the restriction patterns of the chromosomal deoxyribonucleic acid caused by the presence of the abnormal Philadelphia chromosome.
Claim 22
Dependent Claim
24 words
A 1.2 kb fragment of the deoxyribonucleic acid molecule of claim 18 obtained by cleavage with the restriction enzymes Hind III and Bgl II.
Claim 23
Dependent Claim
10 words
A fragment of claim 21 labelled with a detectable marker.
Claim 24
Dependent Claim
136 words
A method of diagnosing chronic myelocytic leukemia in a human subject which is associated with an abnormal Philadelphia chromosome containing the translocated oncogene c-abl which comprises cleaving total chromosomal deoxyribonucleic acid with restriction enzymes under cleaving conditions, separating the fragments so obtained by gel electrophoresis, denaturing the fragments to obtain single-stranded deoxyribonucleic acid molecules, recovering the denatured single-stranded fragments from the gel, immobilizing them on a solid support, contacting the immobilized single-stranded deoxyribonucleic acid fragments with a 1.2 kb deoxyribonucleic acid probe of claim 22 which includes sites for the incorporation of the chromosome 9 segment containing the oncogene c-abl under conditions permitting hybridization of complementary single-stranded molecules, identifying the hybridized molecules so formed to thereby detect abnormalities in the restriction patterns of the chromosomal deoxyribonucleic acid caused by the presence of the abnormal Philadelphia chromosome.
Claim 25
Dependent Claim
24 words
A 0.6 kb fragment of the deoxyribonucleic acid molecule of claim 18 obtained by cleavage with the restriction enzymes Hind III and Bam HI.
Claim 26
Dependent Claim
10 words
A fragment of claim 24 labelled with a detectable marker.
Claim 27
Dependent Claim
137 words
A method of diagnosing chronic myelocytic leukemia in a human subject which is associated with an abnormal Philadelphia chromosome containing the translocated oncogene c-abl which comprises cleaving total chromosomal deoxyribonucleic acid with restriction enzymes under cleaving conditions, separating the fragments so obtained by gel electrophoresis, denaturing the fragments to obtain single-stranded deoxyribonucleic acid molecules, recovering the denatured single-stranded fragments from the gel, immobilizing them on a solid support, contacting the immobilized single-stranded deoxyribonucleic acid fragments with a 0.6 kb deoxyribonucleic acid probe of claim 25 which includes sites for the incorporation of the chromosome 9 segment containing the oncogene c-abl, under suitable conditions permitting hybridization of complementary single-stranded molecules, identifying the hybridized molecules so formed to thereby detect abnormalities in the restriction patterns of the chromosomal deoxyribonucleic acid caused by the presence of the abnormal Philadelphia chromosome.
References Cited (Backward Citations)
US4490472A
Cited
US4535058A
Cited
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1990-07-20 | 1991-01-19 | 1991-07-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-07-20 | 1995-01-19 | 1995-07-21 | $3760.00 | Window Closed / Lapsed |
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