公开(公告)号：US07288375B2

公开(公告)日：2007-10-30

申请号：US10623036

申请日：2003-07-18

申请人： Willem P. C. Stemmer ,  Andreas Crameri

发明人： Willem P. C. Stemmer ,  Andreas Crameri

IPC分类号： C12Q1/68 ,  C12P21/02 ,  C12P19/34 ,  C12P21/08 ,  C07H21/02

CPC分类号： C40B40/02 ,  C07K14/43595 ,  C07K14/545 ,  C07K16/00 ,  C07K2317/565 ,  C07K2317/622 ,  C12N9/2471 ,  C12N9/86 ,  C12N15/1027 ,  C12N15/1034 ,  C12N15/1037 ,  C12Q1/68 ,  C12Q1/6811 ,  C12Y302/01023 ,  C12Y305/02006 ,  C40B40/08 ,  C40B50/06

摘要： A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

摘要翻译： 描述了随机断裂后DNA重组的方法及其通过体外或体内重组诱变核酸序列的应用。 特别地，描述了用于生产编码突变蛋白的核酸片段或多核苷酸的方法。 本发明还涉及一种反复循环诱变，洗牌和选择的方法，其允许蛋白质的体外或体内的定向分子进化。

公开(公告)号：US06287861B1

公开(公告)日：2001-09-11

申请号：US09133508

申请日：1998-08-12

申请人： Willem P. C. Stemmer ,  Andreas Crameri

发明人： Willem P. C. Stemmer ,  Andreas Crameri

IPC分类号： C12N1500

CPC分类号： C40B40/02 ,  C07K14/43595 ,  C07K14/545 ,  C07K16/00 ,  C07K2317/565 ,  C07K2317/622 ,  C12N9/2471 ,  C12N9/86 ,  C12N15/1027 ,  C12N15/1034 ,  C12N15/1037 ,  C12Q1/68 ,  C12Q1/6811 ,  C12Y302/01023 ,  C12Y305/02006 ,  C40B40/08 ,  C40B50/06

摘要： A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

摘要翻译： 描述了随机断裂后DNA重组的方法及其通过体外或体内重组诱变核酸序列的应用。 特别地，描述了用于生产编码突变蛋白的核酸片段或多核苷酸的方法。 本发明还涉及一种反复循环诱变，洗牌和选择的方法，其允许蛋白质的体外或体内的定向分子进化。

公开(公告)号：US5811238A

公开(公告)日：1998-09-22

申请号：US564955

申请日：1995-11-30

申请人： Willem P. C. Stemmer ,  Andreas Crameri

发明人： Willem P. C. Stemmer ,  Andreas Crameri

IPC分类号： C12N15/09 ,  A61K38/00 ,  A61K39/395 ,  C07K14/435 ,  C07K14/545 ,  C07K16/00 ,  C12N9/38 ,  C12N9/86 ,  C12N15/00 ,  C12N15/10 ,  C12P21/08 ,  C12Q1/68 ,  C40B40/02 ,  G01N33/53 ,  G01N33/566

CPC分类号： C40B40/02 ,  C07K14/43595 ,  C07K14/545 ,  C07K16/00 ,  C12N15/1027 ,  C12N15/1034 ,  C12N15/1037 ,  C12N9/2471 ,  C12N9/86 ,  C12Q1/68 ,  C12Q1/6811 ,  C12Y302/01023 ,  C12Y305/02006 ,  C40B40/08 ,  C40B50/06 ,  C07K2317/565 ,  C07K2317/622

摘要： A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

公开(公告)号：US06379964B1

公开(公告)日：2002-04-30

申请号：US09354922

申请日：1999-07-15

申请人： Stephen del Cardayre ,  Matthew Tobin ,  Willem P. C. Stemmer ,  Jeremy Minshull

发明人： Stephen del Cardayre ,  Matthew Tobin ,  Willem P. C. Stemmer ,  Jeremy Minshull

IPC分类号： C12N1500

CPC分类号： A01H1/00 ,  A01K2217/05 ,  C12N15/1027 ,  C12N15/8271 ,  C12N15/8273 ,  C12N15/8279 ,  C12Q1/6811

摘要： The invention provides methods employing iterative cycles of recombination and selection/screening for evolution of whole cells and organisms toward acquisition of desired properties. Examples of such properties include enhanced recombinogenicity, genome copy number, and capacity for expression and/or secretion of proteins and secondary metabolites.

公开(公告)号：US06436675B1

公开(公告)日：2002-08-20

申请号：US09408393

申请日：1999-09-28

申请人： Mark Welch ,  Jon Ness ,  Claes Gustafsson ,  Willem P. C. Stemmer ,  Jeremy Minshull

发明人： Mark Welch ,  Jon Ness ,  Claes Gustafsson ,  Willem P. C. Stemmer ,  Jeremy Minshull

IPC分类号： C12Q168

CPC分类号： C12N15/1031 ,  B01J2219/00689 ,  C12N15/11

摘要： Methods of providing shuffling libraries that include codon-varied oligonucleotide sequences are described. Codon-varied oligonucleotides are synthesized using trinucleotide or mononucleotide phosphoramidite sequences, and are derived from homologous or non-homologous nucleic acid sequences, or combinations of such sequences. Various methods of recombining codon-varied oligonucleotide sequences to expedite artificial evolution are also described. The present invention additionally relates to various integrated systems that are optionally used to automate these recombination methods.

摘要翻译： 描述了提供包括密码子变化的寡核苷酸序列的改组文库的方法。 使用三核苷酸或单核苷酸亚磷酰胺序列合成密码子多样的寡核苷酸，并衍生自同源或非同源核酸序列或这些序列的组合。 还描述了重组密码子变化的寡核苷酸序列以加速人造进化的各种方法。 本发明还涉及可选地用于使这些重组方法自动化的各种集成系统。

公开(公告)号：US07560622B2

公开(公告)日：2009-07-14

申请号：US09970004

申请日：2001-10-01

申请人： Willem P. C. Stemmer

发明人： Willem P. C. Stemmer

IPC分类号： C12N15/31 ,  C12N15/52 ,  C12N15/82

CPC分类号： C12N15/8213

摘要： The present invention provides methods of producing non-transgenic tissues or other agricultural products from a transgenic plant or other transgenic organism. Non-trausgenic organism parts, for example, are derived from transgenic organisms by expression of sequence-specific DNA cleaving enzymes that excise recombinant polynucleotide constructs from the organism's genome by cleaving the constructs at specific recognition sequences that flank the constructs.

公开(公告)号：US06576467B1

公开(公告)日：2003-06-10

申请号：US09718764

申请日：2000-11-21

申请人： Willem P. C. Stemmer

发明人： Willem P. C. Stemmer

IPC分类号： C12N1500

CPC分类号： C40B40/02 ,  C07K14/43595 ,  C07K14/545 ,  C07K16/00 ,  C07K2317/565 ,  C07K2317/622 ,  C12N9/2471 ,  C12N9/86 ,  C12N15/1027 ,  C12N15/1034 ,  C12N15/1037 ,  C12Q1/68 ,  C12Q1/6811 ,  C12Y302/01023 ,  C12Y305/02006 ,  C40B40/08 ,  C40B50/06

摘要： A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

摘要翻译： 描述了随机断裂后DNA重组的方法及其通过体外或体内重组诱变核酸序列的应用。 特别地，描述了用于生产编码突变蛋白的核酸片段或多核苷酸的方法。 本发明还涉及一种反复循环诱变，洗牌和选择的方法，其允许蛋白质的体外或体内的定向分子进化。

公开(公告)号：US06506603B1

公开(公告)日：2003-01-14

申请号：US09717391

申请日：2000-11-18

申请人： Willem P. C. Stemmer

发明人： Willem P. C. Stemmer

IPC分类号： C12N1500

CPC分类号： C12N15/1058 ,  C07K14/43595 ,  C07K14/545 ,  C07K16/00 ,  C07K2317/565 ,  C07K2317/622 ,  C12N9/2471 ,  C12N9/86 ,  C12N15/1027 ,  C12N15/1037 ,  C12N15/52 ,  C12N15/64 ,  C12Q1/68 ,  C12Q1/6811 ,  C12Y302/01023 ,  C40B40/02 ,  Y10S435/876

摘要： A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

摘要翻译： 描述了随机断裂后DNA重组的方法及其通过体外或体内重组诱变核酸序列的应用。 特别地，描述了用于生产编码突变蛋白的核酸片段或多核苷酸的方法。 本发明还涉及一种反复循环诱变，洗牌和选择的方法，其允许蛋白质的体外或体内的定向分子进化。

公开(公告)号：US06500617B1

公开(公告)日：2002-12-31

申请号：US09296886

申请日：1999-04-22

申请人： Willem P. C. Stemmer ,  Linda A. Castle ,  Takashi Yamamoto

发明人： Willem P. C. Stemmer ,  Linda A. Castle ,  Takashi Yamamoto

IPC分类号： C12Q168

CPC分类号： C12N15/8285 ,  C07K14/325 ,  C07K2319/00 ,  C12N15/8286 ,  Y02A40/162 ,  Y02A40/164

摘要： This invention provides methods of obtaining pest resistance genes that are improved over naturally occurring genes for use in conferring upon plants resistance to pests. The methods involve the use of DNA shuffling of pest resistance genes to produce libraries of recombinant pest resistance genes, which are then screened to identify those that exhibit the improved property or properties of interest.

摘要翻译： 本发明提供了获得害虫抗性基因的方法，该基因比用于赋予植物抗病虫害性的天然存在的基因更好。 该方法涉及使用有害动物抗性基因的DNA改组来产生重组害虫抗性基因的文库，然后对其进行筛选以鉴定那些表现出改善的感兴趣的性质或性质的基因。

公开(公告)号：US06165793A

公开(公告)日：2000-12-26

申请号：US75511

申请日：1998-05-08

申请人： Willem P. C. Stemmer

发明人： Willem P. C. Stemmer

IPC分类号： C07K14/435 ,  C07K14/545 ,  C07K16/00 ,  C12N9/38 ,  C12N9/86 ,  C12N15/10 ,  C12N15/52 ,  C12N15/64 ,  C12Q1/68 ,  C40B40/02 ,  C07H21/02 ,  C07H21/04 ,  C12N15/00

CPC分类号： C12N15/1058 ,  C07K14/43595 ,  C07K14/545 ,  C07K16/00 ,  C07K2317/565 ,  C07K2317/622 ,  C12N9/2471 ,  C12N9/86 ,  C12N15/1027 ,  C12N15/1037 ,  C12N15/52 ,  C12N15/64 ,  C12Q1/68 ,  C12Q1/6811 ,  C12Y302/01023 ,  C40B40/02 ,  Y10S435/876

摘要： A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.