Patent EP0234781A2 successfully refreshed from Google Patents.
EP0234781A2
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EP
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Universal restriction endonuclease
Assignee: Individual
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Filing: 1987-02-05
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Granted: —
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Expires: 2007-02-05
Abstract
Abstract A universal restriction endonuclease for cleaving a target DNA at a predetermined site. The universal restriction endonuclease utilizes a tailored oligodeoxynucleotide adapter in conjunction with a restriction enzyme which cuts DNA at a known distance from its recognition site. The oligodeoxynucleotide adapter mimics the relationship which exists in such enzymes between its recognition site and cut site.
Inventors
- 👤 Waclaw Szybalski
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 18:07
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/16; C12N9/22; C12N15/00; C12N15/09; C12N15/10; C12N9/96
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
24 words
A universal restriction endonuclease which cleaves a target DNA at a predetermined site comprising; (a) an oligodeoxynucleotide adapter; and (b) an appropriate restriction enzyme.
Claim 2
Dependent Claim
24 words
A universal restriction endonuclease which cleaves a target DNA at a predetermined site comprising; (a) an oligodeoxynucleotide adapter; and (b) an appropriate restriction enzyme.
Claim 3
Dependent Claim
17 words
The universal restriction endonuclease of claim 1, wherein the oligodeoxynucleotide adapter and restriction enzyme are covalently bound.
Claim 4
Dependent Claim
28 words
The universal restriction endonuclease of claim 1, wherein the oligodeoxynucleotide adapter mimics the relationship which exists in the restriction enzyme between its recognition site and its cleavage site.
Claim 5
Dependent Claim
44 words
The universal restriction endonuclease of claim 1, wherein the restriction enzyme is a Class II enzyme selected from the group of Bbv I, Bbv II, Bin I, Fok I, Hga I, Hph I, Mbo II, Mnl I, Sfa NI, Tag II or Tth lllII.
Claim 6
Dependent Claim
14 words
The universal restriction endonuclease of claim 4, wherein the restriction enzyme is Fok I.
Claim 7
Dependent Claim
41 words
The universal restriction enzyme of claim 4, wherein the oligodeoxynucleotide adapter comprises a double stranded nucleotide sequence corresponding to the recognition site of the restriction enzyme and a single stranded nucleotide sequence complementary to the predetermined site of the target DNA.
Claim 8
Dependent Claim
30 words
The universal restriction endonuclease of claim 1, wherein the restriction enzyme is a Class III enzyme selected from the group of Eco Pl, Eco P15, Hin eI, or Hin fIII.
Claim 9
Dependent Claim
39 words
The universal restriction enzyme of claim 7, wherein the oligodeoxynucleotide adapter comprises a double-stranded nucleotide sequence corresponding to the recognition site of the restriction enzyme and a single-stranded nucleotide sequence complementary to the predetermined site of the target DNA.
Claim 10
Independent Claim
21 words
A method for cleaving a target DNA at a predetermined site comprising contacting the target DNA with a modified restriction endonuclease.
Claim 11
Independent Claim
21 words
A method for cleaving a target DNA at a predetermined site comprising contacting the target DNA with a modified restriction endonuclease.
Claim 12
Dependent Claim
21 words
The method of claim 9, wherein the modified restriction endonuclease comprises: (a) an oligodeoxynucleotide adapter; and (b) an appropriate restriction enzyme.
Claim 13
Dependent Claim
15 words
The method of claim 10, wherein the oligonucleotide adapter and restriction enzyme are covalently bound.
Claim 14
Dependent Claim
37 words
The method of claim 10, wherein the oligonucleotide adapter comprises a double-stranded nucleotide sequence corresponding to the recognition site of the restriction enzyme and a single-stranded nucleotide sequence complementary to the predetermined site of the target DNA.
Claim 15
Dependent Claim
42 words
The method of claim 10, wherein the contacting of the modified restriction endonuclease with the target DNA comprises (a) complexing the oligodeoxynucleotide adapter to the predetermined site of the target DNA; and (b) adding the appropriate restriction enzyme to the DNA-adapter complex.
Claim 16
Dependent Claim
12 words
The method of claim 13, wherein the target DNA is single stranded.
Claim 17
Dependent Claim
41 words
The method of claim 14, comprising the further step of extending the 3ʹ-OH terminus of the oligodeoxynucleotide adapter along the single-stranded target DNA to form a double-stranded DNA containing the predetermined cut site prior to addition of the appropriate restriction enzyme.
Claim 18
Dependent Claim
27 words
The method of claim 10, wherein the oligodeoxynucleotide adapter is complexed with the restriction enzyme and the contacting comprises complexing the enzyme-adapter complex with the target DNA
Claim 19
Dependent Claim
11 words
The method of claim 16, wherein the target DNA is single-stranded.
Claim 20
Dependent Claim
48 words
The method of claim 17, comprising the further step of extending the 3ʹ-OH terminus of the oligodeoxynucleotide adapter along the single-stranded DNA to form a double-stranded DNA, containing the predetermined cut site which double-stranded DNA is then cut by the appropriate restriction enzyme of the enzyme adapter complex.
References Cited (Backward Citations)
No citation records indexed.
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