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1. 发明申请

WO2009017673A3 METHODS, COMPOSITIONS, AND KITS FOR ONE-STEP DNA CLONING USING DNA TOPOISOMERASE 审中-公开
公开(公告)号：WO2009017673A3
公开(公告)日：2009-02-05
申请号：PCT/US2008/009054
申请日：2008-07-25
申请人： DNA TWOPOINTO INC. , NESS, Jon, E. , MINSHULL, Jeremy, S.
发明人： NESS, Jon, E. , MINSHULL, Jeremy, S.
IPC分类号： C40B50/06 , C12N15/00
摘要： Provided herein are methods, compositions, kits, and vectors useful for molecular cloning of, for example, blunt-ended DNA molec using DNA topoisomerase. In certain embodiments, the methods comprise combining into a mixture a first polynucleotide, wherein first polynucleotide comprises an origin of replication, a selectable marker, two topoisomerase recognition sequences, and two nick agent recognition sequences, wherein each of said two topoisomerase recognition sequences is within 0-50 nucleotides of at least of the nicking agent recognition sequences and wherein each of said two nicking agent recognition sequences is nicked, with a sequence-specific topoisomerase and a second polynucleotide, wherein said second polynucleotide comprises a 5' hydroxyl on ea end of the polynucleotide; and transforming the mixture into a host organism, thereby cloning the second polynucleotide.

2. 发明申请

WO2009017673A2 METHODS, COMPOSITIONS, AND KITS FOR ONE-STEP DNA CLONING USING DNA TOPOISOMERASE 审中-公开
标题翻译：使用DNA拓扑异构酶的一步DNA克隆的方法，组合物和试剂盒
公开(公告)号：WO2009017673A2
公开(公告)日：2009-02-05
申请号：PCT/US2008009054
申请日：2008-07-25
申请人： DNA TWOPOINTO INC , NESS JON E , MINSHULL JEREMY S
发明人： NESS JON E , MINSHULL JEREMY S
IPC分类号： C12P19/34 , C12N9/90
CPC分类号： C12N15/66 , C12N9/90 , C12N15/10 , C12N15/64
摘要： Provided herein are methods, compositions, and kits useful for molecular cloning of, for example, blunt-ended DNA molecules using DNA topoisomerase. In certain embodiments, the methods comprise combining into a mixture a first polynucleotide, wherein said first polynucleotide comprises an origin of replication, a selectable marker, two topoisomerase recognition sequences, and two nicking agent recognition sequences, wherein each of said two topoisomerase recognition sequences is within 0-50 nucleotides of at least one of the nicking agent recognition sequences and wherein each of said two nicking agent recognition sequences is nicked, with a sequence- specific topoisomerase and a second polynucleotide, wherein said second polynucleotide comprises a 5 ' hydroxyl on each end of the polynucleotide; and transforming the mixture into a host organism, thereby cloning the second polynucleotide. Advantageously, in some embodiments, the methods, compositions, and kits does not require the formation or purification of a DNA-protein adduct prior to the addition of the polynucleotide to be cloned. Also provided are vector sequences to facilitate performance of the methods and methods for modifying a vector of interest to render it useful in the methods described herein.
摘要翻译：本文提供了可用于例如使用DNA拓扑异构酶的平端DNA分子的分子克隆的方法，组合物和试剂盒。在某些实施方案中，所述方法包括将第一多核苷酸组合成混合物，其中所述第一多核苷酸包含复制起点，可选择标记，两个拓扑异构酶识别序列和两个切口剂识别序列，其中所述两个拓扑异构酶识别序列中的每一个在至少一个切口剂识别序列的0-50个核苷酸内，并且其中每个所述两个切口识别序列被切口，具有序列特异性拓扑异构酶和第二多核苷酸，其中所述第二多核苷酸在每个上包含5'羟基多核苷酸的末端; 并将混合物转化为宿主生物体，从而克隆第二多核苷酸。有利地，在一些实施方案中，方法，组合物和试剂盒在加入待克隆的多核苷酸之前不需要形成或纯化DNA-蛋白质加合物。还提供了用于促进用于修饰感兴趣的载体以使其在本文所述的方法中有用的方法和方法的载体序列。

3. 发明申请

WO2005115102A3 DESIGN, SYNTHESIS AND ASSEMBLY OF SYNTHETIC NUCLEIC ACIDS 审中-公开
标题翻译：合成核酸的设计，合成和组装
公开(公告)号：WO2005115102A3
公开(公告)日：2009-04-09
申请号：PCT/US2005015593
申请日：2005-05-04
申请人： DNA TWOPOINTO INC , GOVINDARAJAN SRIDHAR , KULIKOV NICOLAY V , MINSHULL JEREMY S , NESS JON E
发明人： GOVINDARAJAN SRIDHAR , KULIKOV NICOLAY V , MINSHULL JEREMY S , NESS JON E
IPC分类号： C07H21/00
CPC分类号： C07H21/00
摘要： Methods of synthesizing oligonucleotides with high coupling efficiency (>99.5%) are provided. Methods for purification of synthetic oligonucleotides are also provided. Instrumentation configurations for oligonucleotide synthesis are also provided. Methods of designing and synthesizing polynucleotides are also provided. Polynucleotide design is optimized for subsequent assembly from shorter oligonucleotides. Modifications of phosphoramidite chemistry to improve the subsequent assembly of polynucleotides are provided. The design process also incorporates codon biases into polynucleotides that favor expression in defined hosts. Design and assembly methods are also provided for the efficient synthesis of sets of polynucleotide variants. Software to automate the design and assembly process is also provided.
摘要翻译：提供了具有高耦合效率（> 99.5％）的寡核苷酸的合成方法。还提供了纯化合成寡核苷酸的方法。也提供寡核苷酸合成的仪器配置。还提供了设计和合成多核苷酸的方法。多核苷酸设计被优化用于从较短的寡核苷酸的后续装配。提供亚磷酰胺化学改性以改进随后的多核苷酸组装。设计过程还将密码子偏差结合到有利于在定义的宿主中表达的多核苷酸。还提供了设计和组装方法以有效合成多组DNA变体组。还提供了自动化设计和组装过程的软件。

4. 发明申请

WO2005115102A2 DESIGN, SYNTHESIS AND ASSEMBLY OF SYNTHETIC NUCLEIC ACIDS 审中-公开
标题翻译：合成核酸的设计，合成和组装
公开(公告)号：WO2005115102A2
公开(公告)日：2005-12-08
申请号：PCT/US2005/015593
申请日：2005-05-04
申请人： DNA TWOPOINTO, INC. , GOVINDARAJAN, Sridhar , KULIKOV, Nicolay, V. , MINSHULL, Jeremy, S. , NESS, Jon, E.
发明人： GOVINDARAJAN, Sridhar , KULIKOV, Nicolay, V. , MINSHULL, Jeremy, S. , NESS, Jon, E.
CPC分类号： C07H21/00
摘要： Methods of synthesizing oligonucleotides with high coupling efficiency (>99.5%) are provided. Methods for purification of synthetic oligonucleotides are also provided. Instrumentation configurations for oligonucleotide synthesis are also provided. Methods of designing and synthesizing polynucleotides are also provided. Polynucleotide design is optimized for subsequent assembly from shorter oligonucleotides. Modifications of phosphoramidite chemistry to improve the subsequent assembly of polynucleotides are provided. The design process also incorporates codon biases into polynucleotides that favor expression in defined hosts. Design and assembly methods are also provided for the efficient synthesis of sets of polynucleotide variants. Software to automate the design and assembly process is also provided.
摘要翻译：提供了具有高耦合效率（> 99.5％）的寡核苷酸的合成方法。还提供了纯化合成寡核苷酸的方法。也提供寡核苷酸合成的仪器配置。还提供了设计和合成多核苷酸的方法。多核苷酸设计被优化用于从较短的寡核苷酸的后续装配。提供亚磷酰胺化学改性以改进随后的多核苷酸组装。设计过程还将密码子偏差结合到有利于在定义的宿主中表达的多核苷酸。还提供了设计和组装方法以有效合成多组DNA变体组。还提供了自动化设计和组装过程的软件。

5. 发明申请

WO2003078583A2 OPTIMIZATION OF CROSSOVER POINTS FOR DIRECTED EVOLUTION 审中-公开
标题翻译：用于指导性演变的优势点优化
公开(公告)号：WO2003078583A2
公开(公告)日：2003-09-25
申请号：PCT/US2003/007610
申请日：2003-03-10
申请人： MAXYGEN, INC. , MUNDORFF, Emily, C. , GOVINDARAJAN, Sridhar , GUSTAFSSON, Claes , MINSHULL, Jeremy, S.
发明人： GOVINDARAJAN, Sridhar , GUSTAFSSON, Claes , MINSHULL, Jeremy, S.
IPC分类号： C12N
CPC分类号： G06F19/22 , C40B50/02 , G06F19/16
摘要： Methods and devices for more efficiently engineering diversity into recombinant polypeptides and/or nucleic acids are provided herein. For example, a variety of methods of selecting and/or assessing potential crossover sites in an amino acid sequence or a nucleotide sequence are provided, as well as the resulting chimeric product sequences. These methods include, e.g., consideration of structural, functional and/or statistical data in the selection and assessment of sequences and crossover sites for use in recombination.
摘要翻译：本文提供了用于更有效地将多样性工程化成重组多肽和/或核酸的方法和装置。例如，提供了选择和/或评估氨基酸序列或核苷酸序列中的潜在交叉位点的各种方法，以及所得嵌合产物序列。这些方法包括例如考虑用于重组的序列和交换位点的选择和评估中的结构，功能和/或统计数据。

6. 发明申请

WO2005115102A8 DESIGN, SYNTHESIS AND ASSEMBLY OF SYNTHETIC NUCLEIC ACIDS 审中-公开
标题翻译：合成核酸的设计，合成和组装
公开(公告)号：WO2005115102A8
公开(公告)日：2006-04-06
申请号：PCT/US2005015593
申请日：2005-05-04
申请人： DNA TWOPOINTO INC , GOVINDARAJAN SRIDHAR , KULIKOV NICOLAY V , MINSHULL JEREMY S , NESS JON E
发明人： GOVINDARAJAN SRIDHAR , KULIKOV NICOLAY V , MINSHULL JEREMY S , NESS JON E
CPC分类号： C07H21/00
摘要： Methods of synthesizing oligonucleotides with high coupling efficiency (>99.5%) are provided. Methods for purification of synthetic oligonucleotides are also provided. Instrumentation configurations for oligonucleotide synthesis are also provided. Methods of designing and synthesizing polynucleotides are also provided. Polynucleotide design is optimized for subsequent assembly from shorter oligonucleotides. Modifications of phosphoramidite chemistry to improve the subsequent assembly of polynucleotides are provided. The design process also incorporates codon biases into polynucleotides that favor expression in defined hosts. Design and assembly methods are also provided for the efficient synthesis of sets of polynucleotide variants. Software to automate the design and assembly process is also provided.
摘要翻译：提供了具有高耦合效率（> 99.5％）的寡核苷酸的合成方法。还提供了纯化合成寡核苷酸的方法。也提供寡核苷酸合成的仪器配置。还提供了设计和合成多核苷酸的方法。多核苷酸设计被优化用于从较短的寡核苷酸的后续装配。提供亚磷酰胺化学改性以改进随后的多核苷酸组装。设计过程还将密码子偏差结合到有利于在定义的宿主中表达的多核苷酸。还提供了设计和组装方法以有效合成多组DNA变体组。还提供了自动化设计和组装过程的软件。

7. 发明申请

WO2004099401A1 VARIANT SUBTILISIN ENZYMES (SUBTILASES) 审中-公开
标题翻译：各种分散体酶（SUBTILASES）
公开(公告)号：WO2004099401A1
公开(公告)日：2004-11-18
申请号：PCT/DK2004/000312
申请日：2004-05-06
申请人： NOVOZYMES A/S , MAXIGEN, INC. , MINNING, Stefan , KNÖTZEL, Jürgen, Carsten, Franz , SØRENSEN, Niels Henrik , NESS, Jon, E. , WELCH, Mark, D. , GIVER, Lorraine, J. , CHERRY, Joel , VEDEL BORCHERT, Torben , MINSHULL, Jeremy, S.
发明人： MINNING, Stefan , KNÖTZEL, Jürgen, Carsten, Franz , SØRENSEN, Niels Henrik , NESS, Jon, E. , WELCH, Mark, D. , GIVER, Lorraine, J. , CHERRY, Joel , VEDEL BORCHERT, Torben , MINSHULL, Jeremy, S.
IPC分类号： C12N9/54
CPC分类号： C11D3/38636 , C11D3/38681 , C12N9/52 , Y02P20/582
摘要： Novel subtilases having an improved wash performance on egg stains. These subtilases are useful exhibiting excellent or improved wash performance on egg stains when used in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions.
摘要翻译：在蛋渍上具有改善的洗涤性能的新型脱水酶。当用于例如蛋白质时，这些脱水酶可用于在蛋渍上表现出优异或改善的洗涤性能。清洁剂或洗涤剂组合物，例如衣物洗涤剂组合物和洗碟组合物，包括自动洗碗组合物。

8. 发明申请

WO2003078583A3 OPTIMIZATION OF CROSSOVER POINTS FOR DIRECTED EVOLUTION 审中-公开
公开(公告)号：WO2003078583A3
公开(公告)日：2003-09-25
申请号：PCT/US2003/007610
申请日：2003-03-10
申请人： MAXYGEN, INC. , MUNDORFF, Emily, C. , GOVINDARAJAN, Sridhar , GUSTAFSSON, Claes , MINSHULL, Jeremy, S.
发明人： GOVINDARAJAN, Sridhar , GUSTAFSSON, Claes , MINSHULL, Jeremy, S.
IPC分类号： G06F17/00
摘要： Methods and devices for more efficiently engineering diversity into recombinant polypeptides and/or nucleic acids are provided herein. For example, a variety of methods of selecting and/or assessing potential crossover sites in an amino acid sequence or a nucleotide sequence are provided, as well as the resulting chimeric product sequences. These methods include, e.g., consideration of structural, functional and/or statistical data in the selection and assessment of sequences and crossover sites for use in recombination.

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