CA2485506A1 - A method for in vitro molecular evolution of protein function - Google Patents
A method for in vitro molecular evolution of protein function Download PDFInfo
- Publication number
- CA2485506A1 CA2485506A1 CA002485506A CA2485506A CA2485506A1 CA 2485506 A1 CA2485506 A1 CA 2485506A1 CA 002485506 A CA002485506 A CA 002485506A CA 2485506 A CA2485506 A CA 2485506A CA 2485506 A1 CA2485506 A1 CA 2485506A1
- Authority
- CA
- Canada
- Prior art keywords
- population
- stranded polynucleotide
- polynucleotide molecules
- digestion
- exonuclease
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/102—Mutagenizing nucleic acids
- C12N15/1027—Mutagenizing nucleic acids by DNA shuffling, e.g. RSR, STEP, RPR
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1058—Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
Abstract
A method for in vitro molecular evolution of protein function The invention provides a method for generating a polynucleotide sequence or population of sequences from parent single-stranded polynucleotide sequences encoding one or more protein motifs, comprising the steps of (a) providing a first populatio n of single-stranded polynucleotide molecules and a second population of singl e- stranded polynucleotide molecules, the first and second populations together constituting plus and minus strands of parent polynucleotide sequences, (b) carrying out a reaction for digesting the first and second populations of single-stranded polynucleotide molecules with an exonuclease to generate corresponding populations of single-stranded polynucleotide fragments, (c) contacting said fragments generated from the plus strands with fragments generated from the minus strands and optionally, adding primer sequences tha t anneal to the 3'and 5'ends of at least one of the parent polynucleotides und er annealing conditions, and (d) amplifying the fragments that anneal to each other to generate at least one polynucleotide sequence encoding one or more protein motifs having altered characteristics as compared to the one or more protein motifs encoded by said parent polynucleotides, wherein, in step (b), at least oneparameter of the reaction used for digestion of the first population of single-stranded polynucleotide molecules is different from the equivalent parameter(s) used in the reaction for digestion of the second population of single-stranded polynucleotide molecules. Preferably, the reaction parameter is selected from exonuclease type, exonuclease concentration, reaction volume, duration of the digestion reaction, temperature of the reaction mixture, pH of the reaction mixture, length of parent single~-stranded polynucleotide sequences, amount of single-stranded polynucleotide molecules and buffer composition of the reaction mixture.</SD OAB>
Claims (35)
1. A method for generating a polynucleotide sequence or population of sequences from parent single-stranded polynucleotide sequences encoding one or more protein motifs, the method comprising the steps of a) providing a first population of single-stranded polynucleotide molecules and a second population of single-stranded polynucleotide molecules, the first and second populations together constituting plus and minus strands of parent polynucleotide sequences;
b) carrying out a reaction for digesting the first and second populations of single-stranded polynucleotide molecules with an exonuclease to generate corresponding populations of single-stranded polynucleotide fragments;
c) contacting said polynucleotide fragments generated from the plus strands with fragments generated from the minus strands; and d) amplifying the fragments that anneal to each other to generate at least one polynucleotide sequence encoding one or more protein motifs having altered characteristics as compared to the one or more protein motifs encoded by said parent polynucleotides.
wherein, in step (b), at least one parameter of the reaction used for digestion of the first population of single-stranded polynucleotide molecules is different from the equivalent parameter(s) used in the reaction for digestion of the second population of single-stranded polynucleotide molecules.
b) carrying out a reaction for digesting the first and second populations of single-stranded polynucleotide molecules with an exonuclease to generate corresponding populations of single-stranded polynucleotide fragments;
c) contacting said polynucleotide fragments generated from the plus strands with fragments generated from the minus strands; and d) amplifying the fragments that anneal to each other to generate at least one polynucleotide sequence encoding one or more protein motifs having altered characteristics as compared to the one or more protein motifs encoded by said parent polynucleotides.
wherein, in step (b), at least one parameter of the reaction used for digestion of the first population of single-stranded polynucleotide molecules is different from the equivalent parameter(s) used in the reaction for digestion of the second population of single-stranded polynucleotide molecules.
2. A method according to Claim 1 wherein the reaction parameter is selected from exonuclease type, exonuclease concentration, reaction volume, duration of the digestion reaction, temperature of the reaction mixture, pH of the reaction mixture, length of parent single-stranded polynucleotide sequences, the amount of single-stranded polynucleotide molecules and the buffer composition of the reaction mixture.
3. A method according to Claim 1 or-2 wherein the exonuclease used for digestion of the first population of single-stranded polynucleotide molecules is different from the exonuclease used for digestion of the second population of single-stranded polynucleotide molecules.
4. A method according to Claim 3 wherein the exonuclease used for digestion of the first population of single-stranded polynucleotide molecules is a 3' exonuclease and the exonuclease used for digestion of the second population of single-stranded polynucleotide molecules is a 5' exonuclease.
5. A method according to any one of the preceding claims wherein the exonuclease concentration used for digestion of the first population of single-stranded polynucleotide molecules is different from the exonuclease concentration used for digestion of the second population of single-stranded polynucleotide molecules.
6. A method according to any one of the preceding claims wherein the reaction volume used for digestion of the first population of single-stranded polynucleotide molecules is different from the reaction volume used for digestion of the second population of single-stranded polynucleotide molecules.
7. A method according to any one of the preceding claims wherein the duration of the digestion reaction used for digestion of the first population of single-stranded polynucleotide molecules is different from the duration of the digestion reaction used for digestion of the second population of single-stranded polynucleotide molecules.
8. A method according to any one of the preceding claims wherein the temperature of the reaction mixture used for digestion of the first population of single-stranded polynucleotide molecules is different from the temperature of the reaction mixture used for digestion of the second population of single-stranded polynucleotide molecules.
9. A method according to any one of the preceding claims wherein the pH of the reaction mixture used for digestion of the first population of single-stranded polynucleotide molecules is different from the pH of the reaction mixture used for digestion of the second population of single-stranded polynucleotide molecules.
10. A method according to any one of the preceding claims wherein the length of the polynucleotides in the first population of single-stranded polynucleotide molecules is different from the length of the polynucleotides in the second population of single-stranded polynucleotide molecules.
11. A method according to any one of the preceding claims wherein the buffer composition of the reaction mixture used for digestion of the first population of single-stranded polynucleotide molecules is different from the buffer composition of the reaction mixture used for digestion of the second population of single-stranded polynucleotide molecules.
12. A method according to any one of the preceding claims wherein the amount of single-stranded polynucleotide molecules in the first population of single-stranded polynucleotide molecules is different from the amount of single-stranded polynucleotide molecules in the second population of single-stranded polynucleotide molecules.
13. A method according to any one of the preceding claims wherein the first population of single-stranded polynucleotide molecules constitutes the plus strands of parent polynucleotide sequences and the second population of single-stranded polynucleotide molecules constitutes the minus strands of parent polynucleotide sequences.
14. A method according to any one of the preceding claims wherein the polynucleotide molecules of step (a) are DNA molecules.
15. A method according to any one of the preceding claims wherein step c) further comprises adding primer sequences that anneal to the 3' and/or 5' ends of at least one of the parent polynucleotides under annealing conditions.
16. A method according to any one of the preceding claims wherein the exonuclease used to digest the first and/or second population of single-stranded polynucleotide molecules is selected from the group consisting of BAL 31, exonuclease I, exonuclease V, exonuclease VII, exonuclease T7 gene 6, bacteriophage lambda exonuclease and exonuclease Rec J f.
17. A method according to any one of the preceding claims wherein a parent polynucleotide sequence or sequences has been subjected to mutagenesis.
18. A method according to any one of the preceding claims wherein one or both of the populations of fragments generated in step b) are subjected to mutagenesis.
19. A method according to Claim 17 or 18 wherein the mutagenesis is error prone PCR.
20. A method according to any one of the preceding claims wherein step b) is carried out to generate populations of single-stranded fragments of varying lengths.
21. A method according to Claim 20 wherein step b) is controlled to generate a population of single-stranded fragments having an average length of more than approximately 50 nucleotides.
22. A method according to any one of the preceding claims further comprising the step of expressing at least one polynucleotide sequence generated in step d) to produce the encoded polypeptide.
23. A method according to Claim 22 further comprising the step of testing the encoded polypeptide for desired characteristics.
24. A method according to any one of the preceding claims wherein the parent polynucleotide sequence encodes an antibody or fragment thereof.
25. A method according to any one of the preceding claims wherein the parent polynucleotide sequence encodes an enzyme.
26. A method according to any one of the preceding claims wherein the parent polynucleotide sequence encodes an antigen.
27. A method for making a polypeptide having desired properties, the method comprising the following steps:
(a) generating variant forms of a parent polynucleotide using a method according to any one of Claims 1 to 26;
(b) expressing the variant polynucleotides produced in step (a) to produce variant polypeptides;
(c) screening the variant polypeptides for desired properties; and (d) selecting a polypeptide having desired properties from the variant polypeptides.
(a) generating variant forms of a parent polynucleotide using a method according to any one of Claims 1 to 26;
(b) expressing the variant polynucleotides produced in step (a) to produce variant polypeptides;
(c) screening the variant polypeptides for desired properties; and (d) selecting a polypeptide having desired properties from the variant polypeptides.
28. A polypeptide obtained by a method according to Claim 27.
29. A pharmaceutical composition comprising a polypeptide according to Claim 28 and a pharmaceutically acceptable carrier.
30. A polypeptide according to Claim 28 for use in medicine.
31. Use of a polypeptide according to Claim 28 in the preparation of a medicament for the treatment, therapy and/or diagnosis of a disease.
32. A process for preparing a pharmaceutical composition which comprises, following the identification of a polynucleotide and/or encoded polypeptide with desired characteristics by a method according to any one of Claims 1 to 26, adding said polynucleotide and/or encoded polypeptide to a pharmaceutically acceptable carrier.
33. A process which comprises, following identification of a polynucleotide and/or encoded polypeptide with desired characteristics by a method according to any one of Claims 1 to 26, use of that polynucleotide and/or encoded polypeptide, in whole or in part, in medicine.
34. A process as claimed in Claim 33 wherein the use in medicine is in the treatment, therapy and/or diagnosis of a disease.
35. A process which comprises, following identification of a polynucleotide with desired characteristics according to a method as claimed in any one of Claims 1 to 26, the use of that polynucleotide in the detection and/or amplification of a target polynucleotide in a sample.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB0211369A GB2388604B (en) | 2002-05-17 | 2002-05-17 | A method for in vitro molecular evolution of protein function |
| GB0211369.4 | 2002-05-17 | ||
| US10/321,195 US7153655B2 (en) | 1998-06-16 | 2002-12-17 | Method for in vitro molecular evolution of protein function involving the use of exonuclease enzyme and two populations of parent polynucleotide sequence |
| US10/321,195 | 2002-12-17 | ||
| PCT/GB2003/002102 WO2003097834A2 (en) | 2002-05-17 | 2003-05-16 | A method for in vitro molecular evolution of protein function |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2485506A1 true CA2485506A1 (en) | 2003-11-27 |
| CA2485506C CA2485506C (en) | 2012-02-28 |
Family
ID=29551433
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2485506A Expired - Fee Related CA2485506C (en) | 2002-05-17 | 2003-05-16 | A method for in vitro molecular evolution of protein function |
Country Status (10)
| Country | Link |
|---|---|
| US (1) | US7262012B2 (en) |
| EP (1) | EP1504098B2 (en) |
| JP (1) | JP4372679B2 (en) |
| AT (1) | ATE356871T1 (en) |
| AU (1) | AU2003230027A1 (en) |
| CA (1) | CA2485506C (en) |
| DE (1) | DE60312507T3 (en) |
| DK (2) | DK1504098T4 (en) |
| ES (1) | ES2285118T5 (en) |
| WO (1) | WO2003097834A2 (en) |
Families Citing this family (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| PT1095059E (en) * | 1998-07-10 | 2006-10-31 | Alligator Bioscience Ab Publ | PROTEIN INHIBITOR OF CHEMIAXIA OF ESTAFILOCOCOS (CHIPS) AND ITS UTILIZATION |
| EP1118663A1 (en) * | 2000-01-07 | 2001-07-25 | Universiteit Utrecht | Nucleic acids encoding chemotaxis inhibitory polypeptides |
| US20020086292A1 (en) | 2000-12-22 | 2002-07-04 | Shigeaki Harayama | Synthesis of hybrid polynucleotide molecules using single-stranded polynucleotide molecules |
| EP1586583A3 (en) * | 2004-04-16 | 2005-11-16 | Alligator Bioscience AB (publ) | Compounds that block C5a complement receptor and their use in therapy |
| GB2432366B (en) * | 2005-11-19 | 2007-11-21 | Alligator Bioscience Ab | A method for in vitro molecular evolution of protein function |
| GB0607798D0 (en) | 2006-04-20 | 2006-05-31 | Alligator Bioscience Ab | Novel polypeptides and use thereof |
| GB0708376D0 (en) | 2007-05-01 | 2007-06-06 | Alligator Bioscience Ab | Novel polypeptides and uses thereof |
| GB0905790D0 (en) | 2009-04-03 | 2009-05-20 | Alligator Bioscience Ab | Novel polypeptides and use thereof |
| GB0905503D0 (en) * | 2009-03-31 | 2009-05-13 | Alligator Bioscience Ab | A method for in vitro molecular evolution of protein function |
| GB0920258D0 (en) | 2009-11-19 | 2010-01-06 | Alligator Bioscience Ab | New medical agents and use thereof |
| GB201019086D0 (en) | 2010-11-11 | 2010-12-29 | Imp Innovations Ltd | Bacterial methods |
| WO2012122135A2 (en) | 2011-03-04 | 2012-09-13 | Evolvemol, Inc. | Apparatus and process for isolating specific physical items within a set of physical items |
| GB201115280D0 (en) | 2011-09-05 | 2011-10-19 | Alligator Bioscience Ab | Antibodies, uses and methods |
| GB201311475D0 (en) | 2013-06-27 | 2013-08-14 | Alligator Bioscience Ab | Polypeptides |
| CN114099671A (en) | 2014-08-12 | 2022-03-01 | 鳄鱼生物科学公司 | Combination therapy with anti-CD 40 antibodies |
| BR112018006251A2 (en) | 2015-09-30 | 2018-10-16 | Janssen Biotech Inc | antagonist antibodies that specifically bind to human cd40 and methods of use |
| CN113892654A (en) * | 2021-10-21 | 2022-01-07 | 长春大学 | Method for preparing digestion-free mung bean protein by using mung bean protein powder as raw material |
Family Cites Families (72)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4888286A (en) | 1984-02-06 | 1989-12-19 | Creative Biomolecules, Inc. | Production of gene and protein analogs through synthetic gene design using double stranded synthetic oligonucleotides |
| US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
| DE3546807C2 (en) | 1985-03-30 | 1991-03-28 | Marc Genf/Geneve Ch Ballivet | |
| US5763192A (en) | 1986-11-20 | 1998-06-09 | Ixsys, Incorporated | Process for obtaining DNA, RNA, peptides, polypeptides, or protein, by recombinant DNA technique |
| US4994368A (en) | 1987-07-23 | 1991-02-19 | Syntex (U.S.A.) Inc. | Amplification method for polynucleotide assays |
| US5043272A (en) | 1989-04-27 | 1991-08-27 | Life Technologies, Incorporated | Amplification of nucleic acid sequences using oligonucleotides of random sequence as primers |
| US5023171A (en) | 1989-08-10 | 1991-06-11 | Mayo Foundation For Medical Education And Research | Method for gene splicing by overlap extension using the polymerase chain reaction |
| US5573907A (en) | 1990-01-26 | 1996-11-12 | Abbott Laboratories | Detecting and amplifying target nucleic acids using exonucleolytic activity |
| JP4251406B2 (en) | 1990-04-05 | 2009-04-08 | クレア，ロベルト | Walk-through mutagenesis |
| GB9015198D0 (en) | 1990-07-10 | 1990-08-29 | Brien Caroline J O | Binding substance |
| US5858725A (en) | 1990-10-10 | 1999-01-12 | Glaxo Wellcome Inc. | Preparation of chimaeric antibodies using the recombinant PCR strategy |
| US5514568A (en) | 1991-04-26 | 1996-05-07 | Eli Lilly And Company | Enzymatic inverse polymerase chain reaction |
| US5512463A (en) | 1991-04-26 | 1996-04-30 | Eli Lilly And Company | Enzymatic inverse polymerase chain reaction library mutagenesis |
| DE69230142T2 (en) | 1991-05-15 | 2000-03-09 | Cambridge Antibody Tech | METHOD FOR PRODUCING SPECIFIC BINDING PAIRS |
| US5858657A (en) | 1992-05-15 | 1999-01-12 | Medical Research Council | Methods for producing members of specific binding pairs |
| US5223408A (en) | 1991-07-11 | 1993-06-29 | Genentech, Inc. | Method for making variant secreted proteins with altered properties |
| US5521291A (en) | 1991-09-30 | 1996-05-28 | Boehringer Ingelheim International, Gmbh | Conjugates for introducing nucleic acid into higher eucaryotic cells |
| US5733731A (en) | 1991-10-16 | 1998-03-31 | Affymax Technologies N.V. | Peptide library and screening method |
| US5270170A (en) | 1991-10-16 | 1993-12-14 | Affymax Technologies N.V. | Peptide library and screening method |
| US5252479A (en) | 1991-11-08 | 1993-10-12 | Research Corporation Technologies, Inc. | Safe vector for gene therapy |
| US5733753A (en) | 1992-12-22 | 1998-03-31 | Novo Nordisk A/S | Amplification of genomic DNA by site specific integration of a selectable marker construct |
| US5491074A (en) | 1993-04-01 | 1996-02-13 | Affymax Technologies Nv | Association peptides |
| SE9304060D0 (en) | 1993-12-06 | 1993-12-06 | Bioinvent Int Ab | Methods to select specific bacteriophages |
| US5928905A (en) | 1995-04-18 | 1999-07-27 | Glaxo Group Limited | End-complementary polymerase reaction |
| US6335160B1 (en) | 1995-02-17 | 2002-01-01 | Maxygen, Inc. | Methods and compositions for polypeptide engineering |
| US5837458A (en) | 1994-02-17 | 1998-11-17 | Maxygen, Inc. | Methods and compositions for cellular and metabolic engineering |
| US5605793A (en) | 1994-02-17 | 1997-02-25 | Affymax Technologies N.V. | Methods for in vitro recombination |
| US6309883B1 (en) | 1994-02-17 | 2001-10-30 | Maxygen, Inc. | Methods and compositions for cellular and metabolic engineering |
| US6117679A (en) | 1994-02-17 | 2000-09-12 | Maxygen, Inc. | Methods for generating polynucleotides having desired characteristics by iterative selection and recombination |
| US6165793A (en) | 1996-03-25 | 2000-12-26 | Maxygen, Inc. | Methods for generating polynucleotides having desired characteristics by iterative selection and recombination |
| US6406855B1 (en) | 1994-02-17 | 2002-06-18 | Maxygen, Inc. | Methods and compositions for polypeptide engineering |
| US5834252A (en) | 1995-04-18 | 1998-11-10 | Glaxo Group Limited | End-complementary polymerase reaction |
| US6395547B1 (en) | 1994-02-17 | 2002-05-28 | Maxygen, Inc. | Methods for generating polynucleotides having desired characteristics by iterative selection and recombination |
| US6238884B1 (en) | 1995-12-07 | 2001-05-29 | Diversa Corporation | End selection in directed evolution |
| US6358709B1 (en) | 1995-12-07 | 2002-03-19 | Diversa Corporation | End selection in directed evolution |
| US6352842B1 (en) | 1995-12-07 | 2002-03-05 | Diversa Corporation | Exonucease-mediated gene assembly in directed evolution |
| US5939250A (en) | 1995-12-07 | 1999-08-17 | Diversa Corporation | Production of enzymes having desired activities by mutagenesis |
| US5830696A (en) | 1996-12-05 | 1998-11-03 | Diversa Corporation | Directed evolution of thermophilic enzymes |
| US5965408A (en) * | 1996-07-09 | 1999-10-12 | Diversa Corporation | Method of DNA reassembly by interrupting synthesis |
| US6171820B1 (en) | 1995-12-07 | 2001-01-09 | Diversa Corporation | Saturation mutagenesis in directed evolution |
| US6361974B1 (en) | 1995-12-07 | 2002-03-26 | Diversa Corporation | Exonuclease-mediated nucleic acid reassembly in directed evolution |
| US6368798B1 (en) | 1995-12-07 | 2002-04-09 | Diversa Corporation | Screening for novel bioactivities |
| US6096548A (en) | 1996-03-25 | 2000-08-01 | Maxygen, Inc. | Method for directing evolution of a virus |
| US5851804A (en) | 1996-05-06 | 1998-12-22 | Apollon, Inc. | Chimeric kanamycin resistance gene |
| US5877001A (en) * | 1996-06-17 | 1999-03-02 | Diverso Corporation | Amidase |
| US5763239A (en) * | 1996-06-18 | 1998-06-09 | Diversa Corporation | Production and use of normalized DNA libraries |
| US5925544A (en) | 1996-11-18 | 1999-07-20 | Novo Nordisk A/S | Method of homologous recombination followed by in vivo selection of DNA amplification |
| CN1208456C (en) | 1996-12-20 | 2005-06-29 | 诺维信公司 | Recombination in vivo |
| IL130635A0 (en) | 1997-01-17 | 2000-06-01 | Maxygen Inc | Evolution of whole cells and organisms by recursive sequence recombination |
| US6326204B1 (en) | 1997-01-17 | 2001-12-04 | Maxygen, Inc. | Evolution of whole cells and organisms by recursive sequence recombination |
| AU6029998A (en) | 1997-01-17 | 1998-08-07 | Regents Of The University Of Minnesota | Dna molecules and protein displaying improved triazine compound degrading ability |
| US6159688A (en) | 1997-03-18 | 2000-12-12 | Novo Nordisk A/S | Methods of producing polynucleotide variants |
| US6159687A (en) | 1997-03-18 | 2000-12-12 | Novo Nordisk A/S | Methods for generating recombined polynucleotides |
| US6291165B1 (en) | 1997-03-18 | 2001-09-18 | Novo Nordisk A/S | Shuffling of heterologous DNA sequences |
| US6153410A (en) | 1997-03-25 | 2000-11-28 | California Institute Of Technology | Recombination of polynucleotide sequences using random or defined primers |
| GB9712512D0 (en) * | 1997-06-16 | 1997-08-20 | Bioinvent Int Ab | A method for in vitro molecular evolution of protein function |
| WO1999021979A1 (en) | 1997-10-28 | 1999-05-06 | Maxygen, Inc. | Human papillomavirus vectors |
| US20010006950A1 (en) * | 1998-02-11 | 2001-07-05 | Juha Punnonen | Genetic vaccine vector engineering |
| AU4441699A (en) | 1998-06-17 | 2000-01-05 | Maxygen, Inc. | Method for producing polynucleotides with desired properties |
| US6365408B1 (en) | 1998-06-19 | 2002-04-02 | Maxygen, Inc. | Methods of evolving a polynucleotides by mutagenesis and recombination |
| US20020059659A1 (en) * | 1998-08-12 | 2002-05-16 | Maxygen, Inc. | DNA shuffling to produce herbicide selective crops |
| WO2000009727A2 (en) * | 1998-08-12 | 2000-02-24 | Maxygen, Inc. | Dna shuffling to produce herbicide selective crops |
| US20020151017A1 (en) * | 1998-11-10 | 2002-10-17 | Willem P. C. Stemmer | Methods for obtaining a polynecleotide encoding a polypeptide having a rubisco activity |
| US6358712B1 (en) | 1999-01-05 | 2002-03-19 | Trustee Of Boston University | Ordered gene assembly |
| US6436675B1 (en) * | 1999-09-28 | 2002-08-20 | Maxygen, Inc. | Use of codon-varied oligonucleotide synthesis for synthetic shuffling |
| US6376246B1 (en) * | 1999-02-05 | 2002-04-23 | Maxygen, Inc. | Oligonucleotide mediated nucleic acid recombination |
| US6368861B1 (en) | 1999-01-19 | 2002-04-09 | Maxygen, Inc. | Oligonucleotide mediated nucleic acid recombination |
| EP1165775A2 (en) * | 1999-03-05 | 2002-01-02 | Maxygen, Inc. | Recombination of insertion modified nucleic acids |
| US6329178B1 (en) | 2000-01-14 | 2001-12-11 | University Of Washington | DNA polymerase mutant having one or more mutations in the active site |
| JP3777158B2 (en) * | 2000-11-10 | 2006-05-24 | アミコージェン・インコーポレイテッド | Method for producing recombinant DNA library using unidirectional single-stranded DNA section |
| AU3457202A (en) * | 2000-12-12 | 2002-06-24 | Bioinvent Int Ab | A method for in vitro molecular evolution of protein function |
| US6958213B2 (en) * | 2000-12-12 | 2005-10-25 | Alligator Bioscience Ab | Method for in vitro molecular evolution of protein function |
-
2003
- 2003-05-16 ES ES03722867T patent/ES2285118T5/en not_active Expired - Lifetime
- 2003-05-16 EP EP03722867A patent/EP1504098B2/en not_active Expired - Lifetime
- 2003-05-16 US US10/514,399 patent/US7262012B2/en not_active Expired - Lifetime
- 2003-05-16 DK DK03722867.3T patent/DK1504098T4/en active
- 2003-05-16 WO PCT/GB2003/002102 patent/WO2003097834A2/en active IP Right Grant
- 2003-05-16 JP JP2004506491A patent/JP4372679B2/en not_active Expired - Fee Related
- 2003-05-16 AU AU2003230027A patent/AU2003230027A1/en not_active Abandoned
- 2003-05-16 DE DE60312507T patent/DE60312507T3/en not_active Expired - Lifetime
- 2003-05-16 CA CA2485506A patent/CA2485506C/en not_active Expired - Fee Related
- 2003-05-16 AT AT03722867T patent/ATE356871T1/en active
-
2004
- 2004-11-23 DK DK200401821A patent/DK200401821A/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| DK200401821A (en) | 2004-11-23 |
| WO2003097834A2 (en) | 2003-11-27 |
| DK1504098T3 (en) | 2007-06-11 |
| US20060166198A1 (en) | 2006-07-27 |
| WO2003097834A3 (en) | 2004-02-19 |
| CA2485506C (en) | 2012-02-28 |
| DE60312507T3 (en) | 2011-08-18 |
| AU2003230027A8 (en) | 2003-12-02 |
| DE60312507T2 (en) | 2007-12-06 |
| EP1504098B1 (en) | 2007-03-14 |
| ES2285118T3 (en) | 2007-11-16 |
| JP2005525820A (en) | 2005-09-02 |
| EP1504098A2 (en) | 2005-02-09 |
| AU2003230027A1 (en) | 2003-12-02 |
| ES2285118T5 (en) | 2012-09-21 |
| US7262012B2 (en) | 2007-08-28 |
| EP1504098B2 (en) | 2011-02-09 |
| JP4372679B2 (en) | 2009-11-25 |
| DK1504098T4 (en) | 2011-05-23 |
| ATE356871T1 (en) | 2007-04-15 |
| DE60312507D1 (en) | 2007-04-26 |
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