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31. 发明授权

US6117679A Methods for generating polynucleotides having desired characteristics by iterative selection and recombination 失效
公开(公告)号：US6117679A
公开(公告)日：2000-09-12
申请号：US621859
申请日：1996-03-25
申请人： Willem P. C. Stemmer
发明人： Willem P. C. Stemmer
IPC分类号： C12N15/09 , C07K14/435 , C07K14/46 , C07K14/545 , C07K16/00 , C12N9/38 , C12N9/86 , C12N15/10 , C12N15/52 , C12N15/64 , C12Q1/68 , C40B40/02 , H01L29/772 , H01L29/78 , H01L29/808 , H01L39/22 , C12N15/00 , C07H21/02 , C07H21/04
CPC分类号： C40B40/02 , C07K14/43595 , C07K14/545 , C07K16/00 , C12N15/1027 , C12N15/1034 , C12N15/1037 , C12N15/1058 , C12N15/52 , C12N15/64 , C12N9/2471 , C12N9/86 , C12Q1/68 , C12Q1/6811 , C12Y302/01023 , C12Y305/02006 , H01L39/228 , 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.

32. 发明授权

US5830721A DNA mutagenesis by random fragmentation and reassembly 失效
标题翻译：通过随机碎裂和重新组装进行DNA诱变
公开(公告)号：US5830721A
公开(公告)日：1998-11-03
申请号：US537874
申请日：1996-03-04
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 , C12P19/34
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 generating libraries of displayed peptides and/or antibodies (Abs) suitable for affinity interaction screening or phenotypic screening comprising: (i) obtaining selected library members comprising a displayed peptide and/or Ab and the corressponding polynucleotide (PN), or copies of it, (ii) pooling and fragmenting the PN, or copies of it, to form fragments, (iii) performing PCR amplification and thereby homologously recombining the fragments to form a shuffled pool of recombined PNs, which are not present in the selected library of (i).
摘要翻译： PCT No.PCT / US95 / 02126 Sec。 371日期：1996年3月4日 102（e）日期1996年3月4日PCT提交1995年2月17日PCT公布。 WO95 / 22625 PCT出版物日期1995年8月24日一种用于产生适用于亲和力相互作用筛选或表型筛选的显示肽和/或抗体（Ab）的文库的方法，包括：（i）获得包含显示的肽和/或Ab的选定的文库成员和应答的多核苷酸（ PN）或其拷贝，（ii）合并和分离PN或其拷贝以形成片段，（iii）进行PCR扩增，从而同源重组片段以形成重组PN的混洗池，其不是存在于（i）的选定库中。

33. 发明授权

US5512463A Enzymatic inverse polymerase chain reaction library mutagenesis 失效
标题翻译：酶反向聚合酶链反应库诱变
公开(公告)号：US5512463A
公开(公告)日：1996-04-30
申请号：US252057
申请日：1994-06-01
申请人： Willem P. C. Stemmer
发明人： Willem P. C. Stemmer
IPC分类号： C12N15/10 , C12Q1/68 , C12P19/34 , C07H21/04 , C12P21/00
CPC分类号： C12N15/1093 , C12N15/102 , C12Q1/6844 , C12Q1/6853 , C12Q1/6858
摘要： This invention discloses a method for generating a recombinant library by introducing one or more changes within a predetermined region of double-stranded nucleic acid, comprising providing a first primer population and a second primer population, each of the populations having a variable base composition at known positions along the primers, the primers incorporating a class IIS restriction enzyme recognition sequence, being capable of directing change in the nucleic acid sequence and being substantially complementary to the double stranded nucleic acid to permit hybridization thereto. The method additionally comprises hybridizing the first and second primer populations to opposite strands of the double stranded nucleic acid to form a first pair of primer-templates oriented in opposite directions, performing enzymatic inverse polymerase chain reaction to generate at least one linear copy of the double stranded nucleic acid incorporating the change directed by the primers, cutting the double stranded nucleic acid copy with a class IIS restriction enzyme to form a restricted linear nucleic acid molecule containing the change, joining termini of the restricted linear nucleic acid molecule to produce double-stranded circular nucleic acid and introducing the nucleic acid into compatible host cells. A method is additionally provided for generating a recombinant library using wobble-base mutagenesis.
摘要翻译：本发明公开了一种通过在双链核酸的预定区域内引入一个或多个改变来产生重组文库的方法，其包括提供第一引物群和第二引物群，每个群体具有已知的可变碱基组成沿着引物的位置，引入了IIS限制酶识别序列的引物，能够引导核酸序列的变化并且与双链核酸基本上互补以允许与其进行杂交。该方法另外包括将第一和第二引物群体与双链核酸的相反链杂交，以形成相反方向取向的第一对引物 - 模板，进行酶反向聚合酶链反应以产生双重核酸的至少一个线性拷贝引入由引物引导的变化的双链核酸，用IIS限制酶切割双链核酸拷贝以形成含有限制性线性核酸分子的变化的连接末端以产生双链的限制性线性核酸分子循环核酸并将核酸引入相容的宿主细胞中。另外提供了使用摆动基因突变产生重组文库的方法。

34. 发明授权

US5338665A Peptide library and screening method 失效
标题翻译：肽库和筛选方法
公开(公告)号：US5338665A
公开(公告)日：1994-08-16
申请号：US963321
申请日：1992-10-15
申请人： Peter J. Schatz , Willem P. C. Stemmer
发明人： Peter J. Schatz , Willem P. C. Stemmer
IPC分类号： C12N1/21 , C12N15/10 , C12Q1/68 , G01N33/68 , C12N15/63 , C12N15/72
CPC分类号： G01N33/68 , C12N15/1075
摘要： A random peptide library constructed by transforming host cells with a collection of recombinant vectors that encode a fusion protein comprised of a DNA binding protein and a random peptide and also contain a binding site for the DNA binding protein can be used to screen for novel ligands. The screening method results in the formation of a complex comprising the fusion protein bound to a receptor through the random peptide ligand and to the recombinant DNA vector through the DNA binding protein.
摘要翻译：通过用编码由DNA结合蛋白和随机肽组成的融合蛋白的重组载体的集合转化宿主细胞构建的随机肽文库，并且还含有DNA结合蛋白的结合位点，可用于筛选新配体。筛选方法导致通过DNA结合蛋白形成包含通过随机肽配体与受体结合的融合蛋白和重组DNA载体的复合物。

35. 发明授权

US08058001B2 Oligonucleotide mediated nucleic acid recombination 有权
标题翻译：寡核苷酸介导的核酸重组
公开(公告)号：US08058001B2
公开(公告)日：2011-11-15
申请号：US12557829
申请日：2009-09-11
申请人： Andreas Crameri , Willem P. C. Stemmer , Jeremy Minshull , Steven H. Bass , Mark Welch , John E. Ness , Claes Gustafsson , Phillip A. Patten
发明人： Andreas Crameri , Willem P. C. Stemmer , Jeremy Minshull , Steven H. Bass , Mark Welch , John E. Ness , Claes Gustafsson , Phillip A. Patten
IPC分类号： C12Q1/68 , C12N15/00 , C07H21/00 , C07H21/02 , C07H21/04
CPC分类号： C12P21/06
摘要： Methods of recombining nucleic acids, including homologous nucleic acids, are provided. Families of gene shuffling oligonucleotides and their use in recombination procedures, as well as polymerase and ligase mediated recombination methods are also provided.
摘要翻译：提供重组核酸的方法，包括同源核酸。还提供了基因改组寡核苷酸家族及其在重组过程中的应用，以及聚合酶和连接酶介导的重组方法。

36. 发明授权

US08014961B2 Integrated systems and methods for diversity generation and screening 有权
标题翻译：用于多样化生成和筛选的综合系统和方法
公开(公告)号：US08014961B2
公开(公告)日：2011-09-06
申请号：US11677505
申请日：2007-02-21
申请人： Steven H. Bass , Simon Christopher Davis , Phillip A. Patten , Matthew Tobin , Jeremy S. Minshull , Mark Welch , Claus Gustafsson , Brian Carr , Stephan J. Jenne , Sun Ai Raillard , Andreas Crameri , Willem P. C. Stemmer , Robin Emig , Pascal Longchamp , Stanley Goldman , Lorraine J. Giver , Joseph A. Affholter
发明人： Steven H. Bass , Simon Christopher Davis , Phillip A. Patten , Matthew Tobin , Jeremy S. Minshull , Mark Welch , Claus Gustafsson , Brian Carr , Stephan J. Jenne , Sun Ai Raillard , Andreas Crameri , Willem P. C. Stemmer , Robin Emig , Pascal Longchamp , Stanley Goldman , Lorraine J. Giver , Joseph A. Affholter
IPC分类号： G01N33/48
CPC分类号： C12N15/1031 , B01J19/0046 , B01J2219/00378 , B01J2219/00585 , B01J2219/00659 , B01J2219/00675 , B01J2219/00689 , B01J2219/00695 , B01J2219/007 , B01J2219/00722 , B82Y30/00 , C12N15/1027 , C40B60/04 , G06F19/18 , Y02A90/22
摘要： Integrated systems and methods for diversity generation and screening are provided. The systems use common fluid and array handling components to provide nucleic acid diversification, transcription, translation, product screening and subsequent diversification reactions.
摘要翻译：提供了用于多样化生成和筛选的集成系统和方法。该系统使用普通液体和阵列处理组分来提供核酸多样化，转录，翻译，产物筛选和随后的多样化反应。

37. 发明授权

US07981614B2 Methods for generating polynucleotides having desired characteristics by iterative selection and recombination 失效
标题翻译：通过迭代选择和重组产生具有所需特征的多核苷酸的方法
公开(公告)号：US07981614B2
公开(公告)日：2011-07-19
申请号：US12557519
申请日：2009-09-11
申请人： Willem P. C. Stemmer , Andreas Crameri
发明人： Willem P. C. Stemmer , Andreas Crameri
IPC分类号： C12Q1/68 , C12N15/00 , 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。

38. 发明授权

US07795030B2 Methods and compositions for cellular and metabolic engineering 有权
标题翻译：细胞和代谢工程的方法和组成
公开(公告)号：US07795030B2
公开(公告)日：2010-09-14
申请号：US12011767
申请日：2008-01-28
申请人： Jeremy Minshull , Willem P. C. Stemmer
发明人： Jeremy Minshull , Willem P. C. Stemmer
IPC分类号： C12N15/00 , C12Q1/68 , C07H21/02 , C07H21/04
CPC分类号： C12N15/1027 , C12N15/1058
摘要： The present invention is generally directed to the evolution of new metabolic pathways and the enhancement of bioprocessing through a process herein termed recursive sequence recombination. Recursive sequence recombination entails performing iterative cycles of recombination and screening or selection to “evolve” individual genes, whole plasmids or viruses, multigene clusters, or even whole genomes. Such techniques do not require the extensive analysis and computation required by conventional methods for metabolic engineering.
摘要翻译：本发明一般涉及新代谢途径的演变以及通过本文中称为递归序列重组的方法增强生物处理。递归序列重组需要执行重组和筛选或选择的“进化”个体基因，整个质粒或病毒，多基因簇或甚至全基因组的迭代循环。这些技术不需要传统的代谢工程方法所需的广泛的分析和计算。

39. 发明授权

US07620502B2 Methods for identifying sets of oligonucleotides for use in an in vitro recombination procedure 有权
标题翻译：用于鉴定用于体外重组程序的寡核苷酸组的方法
公开(公告)号：US07620502B2
公开(公告)日：2009-11-17
申请号：US11339090
申请日：2006-01-24
申请人： Sergey A. Selifonov , Willem P. C. Stemmer , Claes Gustafsson , Matthew Tobin , Stephen del Cardayre , Phillip A. Patten , Jeremy Minshull , Lorraine J. Giver
发明人： Sergey A. Selifonov , Willem P. C. Stemmer , Claes Gustafsson , Matthew Tobin , Stephen del Cardayre , Phillip A. Patten , Jeremy Minshull , Lorraine J. Giver
IPC分类号： C01N33/48 , C12Q1/00 , C12Q1/48 , C12Q1/68 , C07H21/02 , C07H21/04
CPC分类号： G06F19/16 , B01J2219/007 , C40B50/02 , G06F19/14 , G06F19/18 , G06F19/22 , G06F19/24
摘要： In silico nucleic acid recombination methods, related integrated systems utilizing genetic operators are used to identify sets of oligonucleotides for in vitro recombination.
摘要翻译：在计算机核酸重组方法中，利用遗传算子的相关综合系统用于鉴定用于体外重组的寡核苷酸组。

40. 发明授权

US07462469B2 Integrated system for diversity generation and screening 失效
标题翻译：用于多样化生成和筛选的综合系统
公开(公告)号：US07462469B2
公开(公告)日：2008-12-09
申请号：US10154936
申请日：2002-05-23
申请人： Steven H. Bass , S. Christopher Davis , Phillip A. Patten , Matthew Tobin , Jeremy Minshull , Mark Welch , Claes Gustafsson , Brian Carr , Stephane Jenne , Sun Ai Raillard , Andreas Crameri , Willem P. C. Stemmer , Robin Emig , Pascal Longchamp , Stanley Goldman , Lorraine J. Giver , Joseph A. Affholter
发明人： Steven H. Bass , S. Christopher Davis , Phillip A. Patten , Matthew Tobin , Jeremy Minshull , Mark Welch , Claes Gustafsson , Brian Carr , Stephane Jenne , Sun Ai Raillard , Andreas Crameri , Willem P. C. Stemmer , Robin Emig , Pascal Longchamp , Stanley Goldman , Lorraine J. Giver , Joseph A. Affholter
IPC分类号： C12P19/34 , C12Q1/68
CPC分类号： C12N15/1031 , B01J19/0046 , B01J2219/00378 , B01J2219/00585 , B01J2219/00659 , B01J2219/00675 , B01J2219/00689 , B01J2219/00695 , B01J2219/007 , B01J2219/00722 , B82Y30/00 , C12N15/1027 , C40B60/04 , G06F19/18 , Y02A90/22
摘要： Integrated systems and methods for diversity generation and screening are provided. The systems use common fluid and array handling components to provide nucleic acid diversification, transcription, translation, product screening and subsequent diversification reactions.
摘要翻译：提供了用于多样化生成和筛选的集成系统和方法。该系统使用普通液体和阵列处理组分来提供核酸多样化，转录，翻译，产物筛选和随后的多样化反应。

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