Patent US5546279A successfully refreshed from Google Patents.
US4667017A
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Method for producing an active protein
Assignee: Asahi Kasei Corp
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Filing: 1984-08-15
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Granted: 1987-05-19
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Expires: 2004-08-15
Abstract
Abstract An active protein can be easily, safely produced by a method comprising providing a first peptide fragment having a first amino acid sequence corresponding to part of an active protein and a second peptide fragment having a second amino acid sequence corresponding to the remaining part of the active protein, at least one of said first peptide fragment and said second peptide fragment being one which has been obtained by recombinant DNA technique or has been obtained by a method comprising producing a predetermined peptide fragment by recombinant DNA technique and deleting from or adding to said predetermined peptide fragment at its N-terminus at least one amino acid residue; and linking said first peptide fragment at its C-terminus to said second peptide fragment at its N-terminus. The method of the present invention may be practiced, with further advantages, by predeterming said first peptide fragment and said second peptide fragment so that a first occurring methionine residue subsequent to the N-terminal amino acid residue of the active protein constitutes the N-terminal amino acid of the amino acid sequence of said second peptide fragment, or so that an amino acid residue positioned near the first occurring methionine residue subsequent to the N-terminal amino acid residue of the desired protein on the side of the N-terminus of the desired protein constitutes the N-terminal amino acid residue of said second peptide fragment.
Inventors
- 👤 Torao Ishida
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 00:01
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6456; C12N9/6459; C07; C07K; C07K1/00; C07K1/107; C07K1/1072; C07K1/1075; C07K14/00; C07K14/435; C07K14/52; C07K14/54; C07K14/55; C07K14/555; C07K14/57; C12Y; C12Y304/00; C12Y304/21; C12Y304/21069; Y; Y02; Y02P; Y02P20/00; Y02P20/50; Y02P20/55; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/14; Y10S930/141; Y10S930/142
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
404 words
A method for producing an active protein having at least methionine residue, arginine residue and lysine residue in its amino acid sequence at an intermediate portion between the N-terminal amino acid residue and the C-terminal amino acid residue of the active protein but not having a methionine residue as the N-terminal amino acid residue, which comprises: (1) providing: (a) a first peptide fragment having a first amino acid sequence corresponding to part of an active protein, said first amino acid sequence containing the N-terminal amino acid residue of the active protein, and (b) a second peptide fragment having a second amino acid sequence corresponding to the remaining part of the active protein, said second amino acid sequence containing the C-terminal amino acid residue of the active protein, at least one of said first peptide fragment and said second peptide fragment being one which has been obtained by a method using recombinant DNA technique, provided that in the case where said first peptide fragment is one which has been obtained by a method using recombinant DNA technique, said first peptide fragment is one which has been obtained by a method comprising producing a first predetermined peptide fragment by means of recombinant DNA technique and deleting from said first predetermined peptide fragment at its N-terminus at least one amino acid residue, and that in the case where said second peptide fragment is one which has been obtained by a method using recombinant DNA technique, said second peptide fragment is one which has been obtained by means of recombinant DNA technique or has been obtained by a method comprising producing a second predetermined peptide fragment by means of recombinant DNA technique and adding to said second predetermined peptide fragment at its N-terminus at least one amino acid residue, said first predetermined peptide fragment being a methionyl peptide containing the N-terminal amino acid residue of the active protein but not containing the peptide residue having an amino acid sequence of from the first occurring methionine residue subsequent to said N-terminal amino acid residue to the C-terminal amino acid residue of said active protein, said second predetermined peptide fragment being a peptide having an amino acid sequence of from the first occurring methionine residue subsequent to the N-terminal amino acid residue to the C-terminal amino acid residue of the active protein; and (2) linking said first peptide fragment at its C-terminus to said second peptide fragment at its N-terminus.
Claim 2
Independent Claim
404 words
A method for producing an active protein having at least methionine residue, arginine residue and lysine residue in its amino acid sequence at an intermediate portion between the N-terminal amino acid residue and the C-terminal amino acid residue of the active protein but not having a methionine residue as the N-terminal amino acid residue, which comprises: (1) providing: (a) a first peptide fragment having a first amino acid sequence corresponding to part of an active protein, said first amino acid sequence containing the N-terminal amino acid residue of the active protein, and (b) a second peptide fragment having a second amino acid sequence corresponding to the remaining part of the active protein, said second amino acid sequence containing the C-terminal amino acid residue of the active protein, at least one of said first peptide fragment and said second peptide fragment being one which has been obtained by a method using recombinant DNA technique, provided that in the case where said first peptide fragment is one which has been obtained by a method using recombinant DNA technique, said first peptide fragment is one which has been obtained by a method comprising producing a first predetermined peptide fragment by means of recombinant DNA technique and deleting from said first predetermined peptide fragment at its N-terminus at least one amino acid residue, and that in the case where said second peptide fragment is one which has been obtained by a method using recombinant DNA technique, said second peptide fragment is one which has been obtained by means of recombinant DNA technique or has been obtained by a method comprising producing a second predetermined peptide fragment by means of recombinant DNA technique and adding to said second predetermined peptide fragment at its N-terminus at least one amino acid residue, said first predetermined peptide fragment being a methionyl peptide containing the N-terminal amino acid residue of the active protein but not containing the peptide residue having an amino acid sequence of from the first occurring methionine residue subsequent to said N-terminal amino acid residue to the C-terminal amino acid residue of said active protein, said second predetermined peptide fragment being a peptide having an amino acid sequence of from the first occurring methionine residue subsequent to the N-terminal amino acid residue to the C-terminal amino acid residue of the active protein; and (2) linking said first peptide fragment at its C-terminus to said second peptide fragment at its N-terminus.
Claim 3
Dependent Claim
87 words
A method according to claim 1, wherein said first peptide fragment and said second peptide fragment are predetermined so that an amino acid residue attached to the C-terminus of said first peptide fragment is reactive with an amino acid residue attached to the N-terminus of said second peptide fragment but the reaction between the amino acid residue attached to the C-terminus of said first peptide fragment and the amino acid residue attached to the N-terminus of said second peptide fragment is not accompanied by a side reaction.
Claim 4
Dependent Claim
14 words
A method according to claim 2, wherein said side reaction is a racemization reaction.
Claim 5
Dependent Claim
52 words
A method according to claim 1, wherein said first peptide fragment and said second peptide fragment are predetermined so that a first occurring methionine residue subsequent to the N-terminal amino acid residue of the active protein constitutes the N-terminal amino acid residue of the amino acid sequence of said second peptide fragment.
Claim 6
Dependent Claim
79 words
A method according to claim 4, wherein the N-terminal amino acid residue of said first peptide fragment is an amino acid residue other than a methionine residue and said first peptide fragment is one which has been obtained by a method comprising producing a first predetermined peptide fragment having a methionine residue as the N-terminal amino acid residue by recombinant DNA technique and deleting said methionine residue as the N-terminal amino acid residue from said first predetermined peptide fragment.
Claim 7
Dependent Claim
18 words
A method according to claim 4, wherein said second peptide fragment is one obtained by recombinant DNA technique.
Claim 8
Dependent Claim
90 words
A method according to claim 1, wherein said first peptide fragment and said second peptide fragment are predetermined so that an amino acid residue positioned near the first occurring methionine residue subsequent to the N-terminal amino acid residue of the active protein on the side of the N-terminus of the active protein constitutes the N-terminal amino acid residue of said second peptide fragment and so that the C-terminal amino acid residue of said first peptide fragment has high reactivity with the N-terminal amino acid residue of said second peptide fragment.
Claim 9
Dependent Claim
75 words
A method according to claim 7, wherein said second peptide fragment is one which has been prepared by a method comprising producing by recombinant DNA technique a second predetermined peptide fragment of which the N-terminal amino acid residue is said first occurring methionine residue subsequent to the N-terminal amino acid residue of the active protein and adding at least one amino acid residue to the N-terminus of said second predetermined peptide fragment by organic synthesis.
Claim 10
Dependent Claim
79 words
A method according to claim 8, wherein the N-terminal amino acid residue of said first peptide fragment is an amino acid residue other than a methionine residue and said first peptide fragment is one which has been obtained by a method comprising producing a first predetermined peptide fragment having a methionine residue as the N-terminal amino acid residue by recombinant DNA technique and deleting said methionine residue as the N-terminal amino acid residue from said first predetermined peptide fragment.
Claim 11
Dependent Claim
28 words
A method according to claim 1, wherein the linking of said first peptide fragment to said second peptide fragment is conducted in the presence of a condensation agent.
Claim 12
Dependent Claim
18 words
A method according to claim 10, wherein said condensation agent is a chemical condensation agent or a protease.
Claim 13
Dependent Claim
62 words
A method according to claim 1, wherein at least one of an amino acid residue attached to the C-terminus of the first peptide fragment and an amino acid residue attached to the N-terminus of the second peptide fragment is converted to an activated form having a reactive group, followed by the linking of said first peptide fragment to said second peptide fragment.
Claim 14
Dependent Claim
19 words
A method according to claim 12, wherein said reactive group is an azido group or a reactive ester group.
Claim 15
Dependent Claim
46 words
A method according to claim 1, wherein one of said first peptide fragment and said second peptide fragment is relatively short in length of the amino acid sequence and the peptide fragment having a relatively short length of amino acid sequence is produced by organic synthesis.
Claim 16
Dependent Claim
18 words
A method according to claim 5, wherein said second peptide fragment is one obtained by recombinant DNA technique.
References Cited (Backward Citations)
No citation records indexed.
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