EP0383620A2
Expired / Lapsed
EP
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Process for making genes encoding random polymers of amino acids
Assignee: Repligen Corp
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Filing: 1990-02-16
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Granted: —
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Expires: 2010-02-16
Abstract
Abstract A method for making a synthetic gene encoding a random polymers of amino-acids but which has predetermined amino acid constituents, comprises the polymerisation of small oligonucleotide duplexes. The novel synthesis can also be used as part of a method for identifying amino-acid polymers with biological and/or immunological activity.
Inventors
- 👤 Kathleen Sue Cook
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 05:49
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C07; C07K; C07K14/00; A; A61; A61K; A61K38/00
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
31 words
A method for making a synthetic gene encoding a random polymer of amino acids where said polymer has predetermined amino acid constituents, said method comprising the polymerization of small oligonucleotide duplexes.
Claim 2
Independent Claim
31 words
A method for making a synthetic gene encoding a random polymer of amino acids where said polymer has predetermined amino acid constituents, said method comprising the polymerization of small oligonucleotide duplexes.
Claim 3
Dependent Claim
30 words
A method, according to claim 1, wherein the length of said synthetic genes is controlled through the use of adaptors specific for the 5′ and 3′ ends of the oligonucleotide.
Claim 4
Dependent Claim
24 words
A method, according to claim 1, wherein said small oligonucleotide duplexes consist of nucleotides where the number of nucleotides is a multiple of 3.
Claim 5
Dependent Claim
28 words
A method, according to claim 3, wherein the size of said small oligonucleotide duplexes is selected from the group consisting of 3-mers, 6-mers, 9-mers, 12-mers, 15-mers, and 18-mers.
Claim 6
Dependent Claim
30 words
A method, according to claim 1, wherein the sticky ends of said duplexes are the same for each duplex so that the duplexes can align and ligate in any order.
Claim 7
Dependent Claim
28 words
A method, according to claim 1, wherein the relative proportion of amino acid constituents in said polymer is determined by the proportion of oligonucleotide duplexes which are incorporated.
Claim 8
Dependent Claim
25 words
A method, according to claim 1, wherein said synthetic gene codes for a polypeptide consisting substantially of alanine, lysine, and either glutamic or aspartic acid.
Claim 9
Dependent Claim
26 words
A method, according to claim 1, wherein said synthetic gene codes for a polypeptide consisting substantially of alanine, lysine, either glutamic or aspartic acid, and tyrosine.
Claim 10
Dependent Claim
29 words
A method, according to claim 1, wherein said synthetic gene codes for a polypeptide consisting substantially of alanine, lysine, either glutamic or aspartic acid, and either tyrosine or tryptophan.
Claim 11
Dependent Claim
20 words
A method, according to claim 1, wherein said polypeptide has a molecular weight between about 5,000 and about 50,000 daltons.
Claim 12
Dependent Claim
25 words
A method, according to claim 1, wherein said synthetic gene codes for a polypeptide which is capable of preventing, arresting, or controlling experimental autoimmune encephalomyelitis.
Claim 13
Dependent Claim
25 words
A method, according to claim 1, wherein said synthetic gene encodes for a polypeptide which is capable of preventing, arresting, or controlling a demyelinating disorder.
Claim 14
Dependent Claim
13 words
A method, according to claim 12, wherein said demyelinating disorder is multiple sclerosis.
Claim 15
Independent Claim
70 words
A method for making polypeptides of random amino acid sequence where said polypeptides have predetermined amino acid constituents, said method comprising the synthesis of genes encoding said random polymers, said synthesis comprising the polymerization of small oligonucleotide duplexes, said method for making polypeptides further comprising the cloning of said synthetic genes into an expression vector and transferring said vector into a bacteria or eukaryotic cell capable of expressing said polypeptide.
Claim 16
Independent Claim
70 words
A method for making polypeptides of random amino acid sequence where said polypeptides have predetermined amino acid constituents, said method comprising the synthesis of genes encoding said random polymers, said synthesis comprising the polymerization of small oligonucleotide duplexes, said method for making polypeptides further comprising the cloning of said synthetic genes into an expression vector and transferring said vector into a bacteria or eukaryotic cell capable of expressing said polypeptide.
Claim 17
Dependent Claim
14 words
A method, according to claim 14, wherein said bacteria is an Escherichia coli .
Claim 18
Independent Claim
102 words
A method for making a fusion polypeptide having one portion comprised of all or part of a heterologous polypeptide and a second portion comprised of a random sequence wherein said random sequence portion has predetermined amino acid constituents, and said method comprises the synthesis of genes encoding said random polymers, said synthesis comprising the polymerization of small oligonucleotide duplexes, said method further comprising the cloning of said synthetic genes into an expression vector adjacent to a DNA sequence encoding all or part of said heterologous polypeptide and transferring said vector into a bacteria or eukaryotic cell capable of expressing said fusion polypeptide.
Claim 19
Independent Claim
102 words
A method for making a fusion polypeptide having one portion comprised of all or part of a heterologous polypeptide and a second portion comprised of a random sequence wherein said random sequence portion has predetermined amino acid constituents, and said method comprises the synthesis of genes encoding said random polymers, said synthesis comprising the polymerization of small oligonucleotide duplexes, said method further comprising the cloning of said synthetic genes into an expression vector adjacent to a DNA sequence encoding all or part of said heterologous polypeptide and transferring said vector into a bacteria or eukaryotic cell capable of expressing said fusion polypeptide.
Claim 20
Independent Claim
143 words
A method for synthesizing and identifying a biologically or immunologically active polypeptide, or mixture of polypeptides, said method comprising the following steps: (a) synthesizing genes encoding random polypeptides by polymerization of small oligonucleotide duplexes; (b) cloning each of said synthetic genes into a vector and transferring said vector into a host capable of expressing said polypeptides; (c) growing said hosts under conditions which permit the formation of recombinant colonies which each express one recombinant gene; (d) testing the polypeptide, or a mixture of the recombinant polypeptides, combined either before or after isolation from said colonies, for evidence of biological and/or immunological activity; (e) where activity is observed for the mixture of polypeptides, generating smaller subsets of that combination and testing each of these subsets for biological activity; and (f) repeating step (e) until the active component(s) or a suitable mixture thereof is obtained.
Claim 21
Independent Claim
143 words
A method for synthesizing and identifying a biologically or immunologically active polypeptide, or mixture of polypeptides, said method comprising the following steps: (a) synthesizing genes encoding random polypeptides by polymerization of small oligonucleotide duplexes; (b) cloning each of said synthetic genes into a vector and transferring said vector into a host capable of expressing said polypeptides; (c) growing said hosts under conditions which permit the formation of recombinant colonies which each express one recombinant gene; (d) testing the polypeptide, or a mixture of the recombinant polypeptides, combined either before or after isolation from said colonies, for evidence of biological and/or immunological activity; (e) where activity is observed for the mixture of polypeptides, generating smaller subsets of that combination and testing each of these subsets for biological activity; and (f) repeating step (e) until the active component(s) or a suitable mixture thereof is obtained.
Claim 22
Dependent Claim
7 words
A synthetic gene which codes for rCOP-1-19.
Claim 23
Dependent Claim
7 words
A synthetic gene which codes for rCOP-1-19.
Claim 24
Dependent Claim
7 words
A synthetic gene which codes for rCOP-1-77.
Claim 25
Dependent Claim
7 words
A synthetic gene which codes for rCOP-1-77.
References Cited (Backward Citations)
No citation records indexed.
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