EP0141484A2
Expired / Lapsed
EP
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Methods of producing hybrid DNA sequences and hybrid polypeptides and DNA sequences produced by them
Assignee: Biogen Inc
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Filing: 1984-06-05
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Granted: —
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Expires: 2004-06-05
Abstract
Abstract @ A method for preparing hybrid DNA sequences characterized by the steps of (a) preparing a DNA fragment, or concatemer thereof, said fragment comprising in sequence one of the parental DNA sequences from which the hybrid DNA sequence is to be derived, an intact replicon such that the DNA fragment may be replicated in a host cell, and the other parental DNA sequence from which the hybrid DNA sequence is to be derived, the two parental DNA sequences having a sequence of at least three, and preferably at least five, nucleotides in common in the location where hybrid DNA sequence formation is sought and the direction of translation of the coding sequences in the two parental sequences being the same relative to each other in the DNA fragment; (b) selecting host cells that have been transfected with the desired hybrid DNA sequence and isolating said hybrid DNA sequence from them. The hybrid DNA sequences of this invention are useful in producing hybrid polypeptides having a variety of uses and biological activities.
Inventors
- 👤 Charles Weissmann
- 👤 Hans Weber
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 18:10
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C07K2319/00; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
144 words
A method for preparing a hybrid DNA sequence comprising the steps of: (a) preparing a DNA fragment, or concatemer thereof, said fragment comprising in sequence one of the parental DNA sequences from which the hybrid DNA sequence is to be derived, an intact replicon such that the DNA fragment may be replicated in a host cell, and the other parental DNA sequence from which the hybrid DNA sequence is to be derived, the two parental DNA sequences having a sequence of at least three nucleotides in common in the location where hybrid DNA sequence formation is sought and the direction of translation of the coding sequences in the two parental sequences being the same relative to each other in the DNA fragment. (b) selecting host cells that have been transfected with the desired hybrid DNA sequence and isolating said hybrid DNA sequence from them.
Claim 2
Independent Claim
144 words
A method for preparing a hybrid DNA sequence comprising the steps of: (a) preparing a DNA fragment, or concatemer thereof, said fragment comprising in sequence one of the parental DNA sequences from which the hybrid DNA sequence is to be derived, an intact replicon such that the DNA fragment may be replicated in a host cell, and the other parental DNA sequence from which the hybrid DNA sequence is to be derived, the two parental DNA sequences having a sequence of at least three nucleotides in common in the location where hybrid DNA sequence formation is sought and the direction of translation of the coding sequences in the two parental sequences being the same relative to each other in the DNA fragment. (b) selecting host cells that have been transfected with the desired hybrid DNA sequence and isolating said hybrid DNA sequence from them.
Claim 3
Dependent Claim
19 words
The method according to claim 1, wherein the parental DNA sequences have homologous sequences of at least 5 nucleotides.
Claim 4
Dependent Claim
19 words
The method according to claim 1, wherein the parental DNA sequences have homologous sequences of at least 8 nucleotides.
Claim 5
Dependent Claim
19 words
The method according to claim 1, wherein the parental DNA sequences have homologous sequences of at least 13 nucleotides.
Claim 6
Independent Claim
22 words
The method of any one of claims 1 to 4, wherein the DNA fragment also comprises at least one antibiotic resistance marker.
Claim 7
Dependent Claim
23 words
The method of claim 5, wherein the DNA fragment comprises a different antibiotic resistance marker associated with each of said parental DNA sequences.
Claim 8
Dependent Claim
52 words
The method of claim 5, wherein the DNA fragment comprises one antibiotic resistance marker and a portion of a different antibiotic resistance marker associated with one of said parental DNA sequences and the portion of said different antibiotic resistance marker that completes it associated with the other of said parental DNA sequences.
Claim 9
Independent Claim
164 words
A method for preparing a hybrid DNA sequence comprising the steps of: (a) preparing a DNA fragment, or concatemer thereof, said fragment comprising in sequence one of the parental DNA segments from which the hybrid DNA sequence is to be derived, an intact replicon such that the fragment may be replicated in a host cell, and the other of the parental DNA sequences from which the hybrid DNA sequence is to be derived, the direction of translation of the coding sequences in the two parental sequences being the same relative to each other in the DNA fragment; (b) forming in one of said parental DNA sequences, a sequence of at least three.nucleotides, said five nucleotide sequence being identical to a DNA sequence in said other parental DNA sequence at the location in said other DNA sequence where said hybrid formation is sought; (c) selecting host cells that have been transfected with the desired hybrid DNA sequence and isolating said hybrid DNA sequences from them.
Claim 10
Independent Claim
164 words
A method for preparing a hybrid DNA sequence comprising the steps of: (a) preparing a DNA fragment, or concatemer thereof, said fragment comprising in sequence one of the parental DNA segments from which the hybrid DNA sequence is to be derived, an intact replicon such that the fragment may be replicated in a host cell, and the other of the parental DNA sequences from which the hybrid DNA sequence is to be derived, the direction of translation of the coding sequences in the two parental sequences being the same relative to each other in the DNA fragment; (b) forming in one of said parental DNA sequences, a sequence of at least three.nucleotides, said five nucleotide sequence being identical to a DNA sequence in said other parental DNA sequence at the location in said other DNA sequence where said hybrid formation is sought; (c) selecting host cells that have been transfected with the desired hybrid DNA sequence and isolating said hybrid DNA sequences from them.
Claim 11
Dependent Claim
23 words
The method of claim 8, wherein a sequence of at least 5 common nucleotides is formed in one of said parental DNA sequences.
Claim 12
Dependent Claim
24 words
The method according to claim 8, wherein a sequence of at least 13 common nucleotides is formed in one of said parental DNA sequences.
Claim 13
Independent Claim
37 words
The method of any one of claims 8 to 10, wherein said step of forming said common nucleotide sequence in one of said parental DNA sequences takes place before the step of preparing said linear DNA fragment.
Claim 14
Independent Claim
22 words
The method of any one of claims 8 to 11, wherein the DNA fragment also comprises at least one antibiotic resistance marker.
Claim 15
Dependent Claim
23 words
The method of claim 12, wherein the DNA fragment comprises a different antibiotic resistance marker associated with each of said parental DNA sequences.
Claim 16
Dependent Claim
52 words
The method of claim 12, wherein the DNA fragment comprises one antibiotic resistance marker and a portion of a different antibiotic resistance marker associated with one of said parental DNA sequences and the portion of said different antibiotic resistance marker that complete it associated with the other of said parental DNA sequences.
Claim 17
Dependent Claim
31 words
The method of claim 1 or 8, wherein said DNA fragment is rendered partially single stranded at its ends and annealed before the step of transfecting said host with the fragment.
Claim 18
Independent Claim
43 words
The method of any one of the preceding claims, wherein said parental DNA sequences are selected from the group consisting of DNA sequences encoding human a interferons, animal a interferons, human p interferons, animal 8 interferons, human y interferons and animal y interferons.
Claim 19
Independent Claim
27 words
The method of any one of claims 1-16, wherein said parental DNA sequences are selected from the group consisting of DNA sequences encoding human and animal lymphokines.
Claim 20
Independent Claim
35 words
The method of any one of claims 1-16, wherein said parental DNA sequences are selected from the group consisting of DNA sequences encoding one or more of the antigens of foot and mouth disease virus.
Claim 21
Dependent Claim
33 words
The method according to claim 6, 7, 13 or 14, wherein said step of selecting said host cells comprises the steps of growing the transformed host cells on agar plates containing both antibiotics.
Claim 22
Dependent Claim
15 words
A hybrid DNA sequence prepared by the method of any one of the preceding claims.
Claim 23
Dependent Claim
78 words
A hybrid DNA sequence comprising at least two connected sequences of nucleotides, each connected DNA sequence being derived from a different parental DNA sequence, the first sequence of nucleotides being connected to the second sequence of nucleotides in the same reading frame so as to maintain constant that reading frame in each sequence through a DNA sequence of at least three nucleotides that was common to both parental DNA sequences at the location where hybrid formation was sought.
Claim 24
Dependent Claim
78 words
A hybrid DNA sequence comprising at least two connected sequences of nucleotides, each connected DNA sequence being derived from a different parental DNA sequence, the first sequence of nucleotides being connected to the second sequence of nucleotides in the same reading frame so as to maintain constant that reading frame in each sequence through a DNA sequence of at least three nucleotides that was common to both parental DNA sequences at the location where hybrid formation was sought.
Claim 25
Dependent Claim
23 words
The hybrid DNA sequence according to claim 21, wherein these are at least five nucleotides that were common to both parental DNA sequences.
Claim 26
Dependent Claim
44 words
The hybrid DNA sequence according to claim 21 or 22, wherein said parental DNA sequences are selected from the group consisting of DNA sequences encoding human a interferons, animal a interferons, human y interferons, animal P interferons, human y interferons and animal y interferons.
Claim 27
Dependent Claim
29 words
The hybrid DNA sequence according to claim 21 or 22, wherein said parental DNA sequences are selected from the group consisting of DNA sequences encoding human and animal lymphokines.
Claim 28
Dependent Claim
37 words
The hybrid DNA sequence according to claim 21 or 22, wherein said parental DNA sequences are selected from the group consisting of DNA sequences encoding one or more of the antigens of foot and mouth disease virus.
Claim 29
Dependent Claim
58 words
A hybrid polypeptide produced by a method comprising the steps of culturing a host transformed with a hybrid DNA sequence selected from the group consisting of hybrid DNA sequences prepared by the methods of any one of claims 1 to 20 and the hybrid DNA sequences of any one of claims 21 to 25 and collecting said polypeptide.
Claim 30
Dependent Claim
65 words
A hybrid polypeptide comprising at least two connected sequences of amino acids, each connected amino acid sequence being derived from a different parental polypeptide, the first sequence of amino acids being connected to the second sequence of amino acids through an amino acid sequence of at least one amino acid that was common to both parental polypeptides at the location where hybrid formation was sought.
Claim 31
Dependent Claim
65 words
A hybrid polypeptide comprising at least two connected sequences of amino acids, each connected amino acid sequence being derived from a different parental polypeptide, the first sequence of amino acids being connected to the second sequence of amino acids through an amino acid sequence of at least one amino acid that was common to both parental polypeptides at the location where hybrid formation was sought.
Claim 32
Dependent Claim
27 words
The hybrid polypeptide according to claim 27, wherein at least two amino acids were common to both parental polypeptides at the location where hybrid formation was sought.
References Cited (Backward Citations)
No citation records indexed.
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