US4935357A
Expired / Lapsed
US
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Universal restriction endonuclease
Assignee: New England Biolabs Inc
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Filing: 1986-02-05
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Granted: 1990-06-19
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Expires: 2006-02-05
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
US
Data Source
google_patents
Last Synchronized
2026-10-29 19:13
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: 14
Claim 1
Independent Claim
76 words
A universal restriction endonuclease composition which cleaves a target DNA at a predetermined site comprising: (a) an oligonucleotide adapter which comprises a double-stranded nucleotide sequence corresponding to the recognition site of the restriction endonuclease and a single-stranded nucleotide sequence complementary to the predetermined site of the target DNA: and (b) a restriction endonuclease; wherein the oligodeoxynucleotide adapter is designed to mimic the relationship which exists between the recognition site and cleavage site of the restriction endonuclease.
Claim 2
Dependent Claim
76 words
A universal restriction endonuclease composition which cleaves a target DNA at a predetermined site comprising: (a) an oligonucleotide adapter which comprises a double-stranded nucleotide sequence corresponding to the recognition site of the restriction endonuclease and a single-stranded nucleotide sequence complementary to the predetermined site of the target DNA: and (b) a restriction endonuclease; wherein the oligodeoxynucleotide adapter is designed to mimic the relationship which exists between the recognition site and cleavage site of the restriction endonuclease.
Claim 3
Dependent Claim
18 words
The universal restriction endonuclease composition of claim 1, wherein the oligodeoxynucleotide adapter and restriction endonuclease are covalently bound.
Claim 4
Dependent Claim
35 words
The universal restriction endonuclease composition of claim 1, wherein the restriction endonuclease is a Class II enzyme selected from the group consisting of BbvI, BbvII, BinI, FokI, HgaI, HphI, MboII, MnlI, SfaNI, TaqII, and Tth111II.
Claim 5
Dependent Claim
14 words
The universal restriction endonuclease composition of claim 3, wherein the restriction endonuclease is FokI.
Claim 6
Dependent Claim
28 words
The universal restriction endonuclease composition of claim 1, wherein the restriction endonuclease is a Class III enzyme selected from the group consisting of EcoP1, EcoP15, HineI, and HinfIII.
Claim 7
Independent Claim
109 words
A method for cleaving a target DNA at a predetermined site comprising contacting the target DNA in an appropriate buffer solution with a universal restriction endonuclease composition in an amount and for a time sufficient to cause the desired cleavage to occur, said universal restriction endonuclease composition comprising: (a) an oligonucleotide adapter which comprises a double-stranded nucleotide sequence corresponding to the recognition site of the restriction endonuclease and a single-stranded nucleotide sequence complementary to the predetermined site of the target DNA; and (b) a restriction endonuclease; wherein the oligodeoxynucleotide adapter is designed to mimic the relationship which exists between the recognition site and cleavage site of the restriction endonuclease.
Claim 8
Independent Claim
109 words
A method for cleaving a target DNA at a predetermined site comprising contacting the target DNA in an appropriate buffer solution with a universal restriction endonuclease composition in an amount and for a time sufficient to cause the desired cleavage to occur, said universal restriction endonuclease composition comprising: (a) an oligonucleotide adapter which comprises a double-stranded nucleotide sequence corresponding to the recognition site of the restriction endonuclease and a single-stranded nucleotide sequence complementary to the predetermined site of the target DNA; and (b) a restriction endonuclease; wherein the oligodeoxynucleotide adapter is designed to mimic the relationship which exists between the recognition site and cleavage site of the restriction endonuclease.
Claim 9
Dependent Claim
15 words
The method of claim 6, wherein the oligonucleotide adapter and restriction endonuclease are covalently bound.
Claim 10
Dependent Claim
43 words
The method of claim 6, wherein the contacting of the universal restriction endonuclease composition with the target DNA comprises: (a) complexing the oligodeoxynucleotide adapter to the predetermined site of the target DNA; and (b) adding the appropriate restriction endonuclease to the DNA-adapter complex.
Claim 11
Dependent Claim
12 words
The method of claim 8, wherein the target DNA is single stranded.
Claim 12
Dependent Claim
41 words
The method of claim 9, comprising the further step of extending the 3'-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 endonuclease.
Claim 13
Dependent Claim
25 words
The method of claim 6, wherein the oligodeoxynucleotide adapter is complexed with the restriction endonuclease and contacting comprises complexing the complex with the target DNA.
Claim 14
Dependent Claim
11 words
The method of claim 11 wherein the target DNA is single-stranded.
References Cited (Backward Citations)
US4581333A
Cited
US4670379A
Cited
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