EP0074808A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Recombinant method and materials
Assignee: University Patents Inc
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Filing: 1982-09-09
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Granted: —
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Expires: 2002-09-09
Abstract
Abstract Specific DNA sequence insertions, deletions and substitutions (i.e., combinations of sequence deletion and inser- tiona) in eukaryotic cell or viral genomes are stably effected through use of selectable DNA sequences comprising a herpesvirus thymidine kinase (tk) gene.
Inventors
- 👤 Bernard Roizman
- 👤 Leonard E. Post
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 15:31
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/87; C12N15/90; C12N15/902
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
119 words
A method for stably effecting the insertion or deletion of a selected DNA sequence at a specific site in a eukaryotic or viral genome, said method comprising: (1) isolating from said genome a linear DNA fragment comprising both (a) the specific site determined for insertion or deletion of selected DNA sequence and (b) flanking DNA sequences normally preceding and following said site; (2) preparing first and second altered genome fragments from the fragment isolated in step (1), (a) said first altered fragment comprising the fragment including a herpesvirus thymidine kinase gene in a position intermediate the ends of said fragment, and (b) said second altered fragment comprising the fragment having said selected DNA sequence inserted therein or deleted therefrom;
Claim 2
Independent Claim
119 words
A method for stably effecting the insertion or deletion of a selected DNA sequence at a specific site in a eukaryotic or viral genome, said method comprising: (1) isolating from said genome a linear DNA fragment comprising both (a) the specific site determined for insertion or deletion of selected DNA sequence and (b) flanking DNA sequences normally preceding and following said site; (2) preparing first and second altered genome fragments from the fragment isolated in step (1), (a) said first altered fragment comprising the fragment including a herpesvirus thymidine kinase gene in a position intermediate the ends of said fragment, and (b) said second altered fragment comprising the fragment having said selected DNA sequence inserted therein or deleted therefrom;
Claim 3
Dependent Claim
35 words
(3) contacting said genome with said first altered fragment under conditions permitting recombination at sites of DNA sequence homology, selecting for a recombinant genome including said thymidine kinase gene, and isolating the recombinant genome; and
Claim 4
Dependent Claim
35 words
(3) contacting said genome with said first altered fragment under conditions permitting recombination at sites of DNA sequence homology, selecting for a recombinant genome including said thymidine kinase gene, and isolating the recombinant genome; and
Claim 5
Dependent Claim
40 words
(4) contacting the recombinant genome isolated in step (3) with said second altered fragment under conditions permitting recombination at sites of DNA sequence homology, selecting for a recombinant genome lacking said thymidine kinase gene, and isolating the recombinant genome product.
Claim 6
Dependent Claim
40 words
(4) contacting the recombinant genome isolated in step (3) with said second altered fragment under conditions permitting recombination at sites of DNA sequence homology, selecting for a recombinant genome lacking said thymidine kinase gene, and isolating the recombinant genome product.
Claim 7
Dependent Claim
13 words
A method according to claim 1 wherein the genome is a viral genome.
Claim 8
Dependent Claim
14 words
A method according to claim 2 wherein the genome is a pathogenic viral genome.
Claim 9
Dependent Claim
19 words
A method according to claim 1 wherein said herpesvirus thymidine kinase gene is of human Herpes Virus 1 origin.
Claim 10
Dependent Claim
31 words
A method according to claim 1 wherein the step of selecting for a recombinant genome including said thymidine kinase gene -involves exposure of the genome to a growth medium including HAT.
Claim 11
Independent Claim
34 words
A method according to claims 1 or 5 wherein the step of selecting for a recombinant genome not including said thymidine kinase gene involves exposure of the genome to a growth medium including AraT.
Claim 12
Dependent Claim
26 words
A method according to claim 1 wherein a deletion of one selected DNA sequence and the in- . sertion of another selected DNA sequence is effected.
Claim 13
Dependent Claim
20 words
A method according to claim 7 wherein the deletion and insertion are at substantially the same site in the genome.
Claim 14
Dependent Claim
18 words
A method according to claim 7 wherein the deletion and insertion are at different sites in the genome.
Claim 15
Dependent Claim
15 words
A method according to claim 9 wherein the DNA sequences inserted and deleted are identical.
Claim 16
Dependent Claim
28 words
A viral genome which is the product of an insertion or deletion of a selected DNA sequence at a specific site according to the method of claim 1.
Claim 17
Dependent Claim
29 words
A eukaryotic cell genome which is the product of an insertion or deletion of a selected DNA sequence at a specific site according to the method of claim 1.
References Cited (Backward Citations)
No citation records indexed.
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