US6492107B1
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for obtaining DNA, RNA, peptides, polypeptides, or protein, by recombinant DNA technique
Assignee: SANTA FE INSTITUTE
•
Filing: 1995-06-05
•
Granted: 2002-12-10
•
Expires: 2015-06-05
Abstract
Abstract The present invention is directed to a process for the production of a peptide, polypeptide, or protein having a predetermined property. In accordance with one embodiment, the process begins by producing by way of synthetic polynucleotide coupling, stochastically generated polynucleotide sequences. A library of expression vectors containing such stochastically generated polynucleotide sequences is formed. Next, host cells containing the vectors are cultured so as to produce peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide sequences. Screening or selection is carried out on such host cells to identify a peptide, polypeptide, or protein produced by the host cells which has the predetermined property. The stochastically generated polynucleotide sequence which encodes the identified peptide, polypeptide, or protein is then isolated and used to produce the peptide, polypeptide, or protein having the predetermined property.
Inventors
- 👤 Stuart Alan Kauffman
- 👤 Marc Ballivet
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 12:38
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1034; C12N15/1093
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
121 words
A process for the production of a peptide, polypeptide, or protein having a predetermined property, comprising the steps of: producing stochastically generated polynucleotide sequences; forming a library of expression vectors comprising said stochastically generated polynucleotide sequences; culturing host cells comprising said library of expression vectors to produce said peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide. sequences; screening or selecting said host cells, to thereby identify a host cell comprising an expression vector encoding a peptide, polypeptide, or protein having the predetermined property; isolating a stochastically generated polynucleotide sequence from said expression vector which encodes the identified peptide, polypeptide, or protein; using the isolated stochastically generated polynucleotide sequence to produce the peptide, polypeptide, or protein having the predetermined property.
Claim 2
Dependent Claim
121 words
A process for the production of a peptide, polypeptide, or protein having a predetermined property, comprising the steps of: producing stochastically generated polynucleotide sequences; forming a library of expression vectors comprising said stochastically generated polynucleotide sequences; culturing host cells comprising said library of expression vectors to produce said peptides, polypeptides, or proteins encoded by the stochastically generated polynucleotide. sequences; screening or selecting said host cells, to thereby identify a host cell comprising an expression vector encoding a peptide, polypeptide, or protein having the predetermined property; isolating a stochastically generated polynucleotide sequence from said expression vector which encodes the identified peptide, polypeptide, or protein; using the isolated stochastically generated polynucleotide sequence to produce the peptide, polypeptide, or protein having the predetermined property.
Claim 3
Dependent Claim
118 words
A process for the production of a peptide, polypeptide, or protein having a predetermined property, comprising the steps of: producing a population of stochastic polynucleotide sequences comprising stochastic synthetic polynucleotide sequences; introducing the population of stochastic polynucleotide sequences thus obtained into host cells to produce transformed host cells; cultivating the transformed host cells comprising said population of stochastic polynucleotide sequences to produce peptides, polypeptides, or proteins encoded by stochastic polynucleotide sequences; screening or selecting a cell from said transformed host cells thereby identifying a cell producing the peptide, polypeptide, or protein having the predetermined property; isolating the cell so identified, and growing the isolated cell under conditions to produce the peptide, polypeptide, or protein having the predetermined property.
Claim 4
Dependent Claim
118 words
A process for the production of a peptide, polypeptide, or protein having a predetermined property, comprising the steps of: producing a population of stochastic polynucleotide sequences comprising stochastic synthetic polynucleotide sequences; introducing the population of stochastic polynucleotide sequences thus obtained into host cells to produce transformed host cells; cultivating the transformed host cells comprising said population of stochastic polynucleotide sequences to produce peptides, polypeptides, or proteins encoded by stochastic polynucleotide sequences; screening or selecting a cell from said transformed host cells thereby identifying a cell producing the peptide, polypeptide, or protein having the predetermined property; isolating the cell so identified, and growing the isolated cell under conditions to produce the peptide, polypeptide, or protein having the predetermined property.
Claim 5
Dependent Claim
76 words
A process for the production of a DNA having a selectable predetermined property, comprising the steps of: producing DNA comprising stochastically generated polynucleotide sequences; forming a library of expression vectors comprising said stochastically generated polynucleotide sequences; culturing host cells comprising expression vectors from said library of expression vectors to express said DNA, and screening or selecting said host cells to identify a host cell comprising an expression vector expressing said DNA having the selectable predetermined property.
Claim 6
Dependent Claim
76 words
A process for the production of a DNA having a selectable predetermined property, comprising the steps of: producing DNA comprising stochastically generated polynucleotide sequences; forming a library of expression vectors comprising said stochastically generated polynucleotide sequences; culturing host cells comprising expression vectors from said library of expression vectors to express said DNA, and screening or selecting said host cells to identify a host cell comprising an expression vector expressing said DNA having the selectable predetermined property.
Claim 7
Dependent Claim
24 words
The process according to claim 3 , wherein said selectable predetermined property comprises the capacity of said DNA to selectively bind to a compound.
Claim 8
Dependent Claim
93 words
A process for the production of a DNA having a selectable predetermined property, comprising the steps of: producing a population of DNA comprising stochastic synthetic polynucleotide sequences; introducing said population of DNA thus obtained into host cells to produce transformed host cells; cultivating the transformed host cells; screening or selecting the transformed host cells to identify a cell producing a DNA having said selectable predetermined property; isolating the cell producing said DNA having said selectable predetermined property, and growing the isolated cell under conditions to produce the DNA having the selectable predetermined property.
Claim 9
Dependent Claim
93 words
A process for the production of a DNA having a selectable predetermined property, comprising the steps of: producing a population of DNA comprising stochastic synthetic polynucleotide sequences; introducing said population of DNA thus obtained into host cells to produce transformed host cells; cultivating the transformed host cells; screening or selecting the transformed host cells to identify a cell producing a DNA having said selectable predetermined property; isolating the cell producing said DNA having said selectable predetermined property, and growing the isolated cell under conditions to produce the DNA having the selectable predetermined property.
Claim 10
Dependent Claim
85 words
A process for the production of a population of DNA comprising stochastic polynucleotide sequences, comprising the steps of: linearizing an expression vector; reacting said linearized expression vector with terminal transferase enzyme in the presence of predetermined ratios of deoxynucleotide-triphosphates of guanine, cytosine, thymidine, and adenine to form a stochastic polynucleotide sequence at each 3′ extremity of said linearized vector; hybridizing said stochastic polynucleotide sequence at a 3′ extremity of said linearized vector, and synthesizing a second strand from said 3′ ends of said hybridized vector.
Claim 11
Dependent Claim
85 words
A process for the production of a population of DNA comprising stochastic polynucleotide sequences, comprising the steps of: linearizing an expression vector; reacting said linearized expression vector with terminal transferase enzyme in the presence of predetermined ratios of deoxynucleotide-triphosphates of guanine, cytosine, thymidine, and adenine to form a stochastic polynucleotide sequence at each 3′ extremity of said linearized vector; hybridizing said stochastic polynucleotide sequence at a 3′ extremity of said linearized vector, and synthesizing a second strand from said 3′ ends of said hybridized vector.
Claim 12
Dependent Claim
47 words
A process for the production of a library of DNA molecules comprising stochastic sequences of polynucleotides, comprising the steps of: producing stochastic sequences of polynucleotides; ligating said stochastic sequences of polynucleotides into expression vectors; transforming competent cells with said ligated expression vectors; and amplifying said transformed cells.
Claim 13
Dependent Claim
47 words
A process for the production of a library of DNA molecules comprising stochastic sequences of polynucleotides, comprising the steps of: producing stochastic sequences of polynucleotides; ligating said stochastic sequences of polynucleotides into expression vectors; transforming competent cells with said ligated expression vectors; and amplifying said transformed cells.
Claim 14
Dependent Claim
72 words
A process for the production of an RNA molecule having a selectable predetermined property, comprising the steps of: producing a population of DNA molecules comprising stochastically generated polynucleotide sequences; forming a library of expression vectors comprising said stochastically generated polynucleotide sequences; culturing host cells comprising said expression vectors forming said library to express RNA from said DNA; screening or selecting said host cells to identify a RNA having the selectable predetermined property.
Claim 15
Dependent Claim
72 words
A process for the production of an RNA molecule having a selectable predetermined property, comprising the steps of: producing a population of DNA molecules comprising stochastically generated polynucleotide sequences; forming a library of expression vectors comprising said stochastically generated polynucleotide sequences; culturing host cells comprising said expression vectors forming said library to express RNA from said DNA; screening or selecting said host cells to identify a RNA having the selectable predetermined property.
Claim 16
Dependent Claim
96 words
A process for the production of an RNA molecule having a selectable predetermined property, comprising the steps of: producing a population of DNA molecules comprising stochastic synthetic polynucleotide sequences, introducing said population of DNA molecules into host cells to produce transformed host cells; cultivating said transformed host cells to produce a population of RNA comprising stochastic polynucleotide sequences; screening or selecting said transformed host cells to identify a cell expressing an RNA molecule having the selectable predetermined property; isolating the identified cell; growing said isolated cell, and producing said RNA molecule having the selectable predetermined property.
Claim 17
Dependent Claim
96 words
A process for the production of an RNA molecule having a selectable predetermined property, comprising the steps of: producing a population of DNA molecules comprising stochastic synthetic polynucleotide sequences, introducing said population of DNA molecules into host cells to produce transformed host cells; cultivating said transformed host cells to produce a population of RNA comprising stochastic polynucleotide sequences; screening or selecting said transformed host cells to identify a cell expressing an RNA molecule having the selectable predetermined property; isolating the identified cell; growing said isolated cell, and producing said RNA molecule having the selectable predetermined property.
Claim 18
Dependent Claim
26 words
The process according to claim 8 or 9 , wherein said selectable predetermined property comprises the capacity of said RNA molecule to selectively bind a compound.
Claim 19
Dependent Claim
68 words
A process for the production of a library of RNA molecules comprising stochastic sequences of polynucleotide, comprising the steps of: producing DNA comprising stochastic polynucleotide sequences; ligating said DNA comprising stochastic polynucleotide sequences into expression vectors capable of expressing said DNA to produce said RNA; transforming competent cells with said ligated expression vectors; amplifying said transformed cells, and expressing said DNA to produce said library of molecule RNA.
Claim 20
Dependent Claim
68 words
A process for the production of a library of RNA molecules comprising stochastic sequences of polynucleotide, comprising the steps of: producing DNA comprising stochastic polynucleotide sequences; ligating said DNA comprising stochastic polynucleotide sequences into expression vectors capable of expressing said DNA to produce said RNA; transforming competent cells with said ligated expression vectors; amplifying said transformed cells, and expressing said DNA to produce said library of molecule RNA.
Claim 21
Dependent Claim
85 words
A process for the production of a population of DNA comprising stochastic polynucleotide sequences, comprising the steps of: linearizing an expression vector; reacting said linearized expression vector with terminal transferase enzyme in the presence of stochastic ratios of deoxynucleotide-triphosphates of guanine, cytosine, thymidine, and adenine to form a stochastic polynucleotide sequence at each 3′ extremity of said linearized vector; hybridizing said stochastic polynucleotide sequence at a 3′ extremity of said linearized vector; and synthesizing a second strand from said 3′ ends of said hybridized vector.
Claim 22
Dependent Claim
85 words
A process for the production of a population of DNA comprising stochastic polynucleotide sequences, comprising the steps of: linearizing an expression vector; reacting said linearized expression vector with terminal transferase enzyme in the presence of stochastic ratios of deoxynucleotide-triphosphates of guanine, cytosine, thymidine, and adenine to form a stochastic polynucleotide sequence at each 3′ extremity of said linearized vector; hybridizing said stochastic polynucleotide sequence at a 3′ extremity of said linearized vector; and synthesizing a second strand from said 3′ ends of said hybridized vector.
References Cited (Backward Citations)
US4366246A
Cited
US4490358A
Cited
US4366264A
Cited
DE3246071A1
Cited
DE3300632A1
Cited
DE3303173A1
Cited
US4634678A
Cited
US4959312A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2005-12-09 | 2006-06-10 | 2006-12-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2009-12-09 | 2010-06-10 | 2010-12-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2013-12-09 | 2014-06-10 | 2014-12-10 | $7700.00 | Window Closed / Lapsed |