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30 条记录 (耗时 0.058 秒)

    Amplification and cloning of single dna molecules using rolling circle amplification
    12.
    发明申请
    Amplification and cloning of single dna molecules using rolling circle amplification 有权
    使用滚圆放大扩增和克隆单个dna分子

    公开(公告)号：US20090176280A1

    公开(公告)日：2009-07-09

    申请号：US11919515

    申请日：2006-05-01

    申请人： Clyde A. Hutchison III ,  Hamilton O. Smith

    发明人： Clyde A. Hutchison III ,  Hamilton O. Smith

    IPC分类号： C12P19/34

    CPC分类号： C12N15/1075 ,  C12N15/64 ,  C12Q1/6848 ,  C12Q2531/119 ,  C12Q2527/143 ,  C12Q2525/204 ,  C12Q2531/125

    摘要： The present invention relates, e.g., to a method for amplifying a small number of copies (e.g. a single copy) of a single-stranded circular DNA molecule (e.g. having a size of about 5-6 kb) by an isothermal rolling circle mechanism, using random or partially random primers and a F29-type DNA polymerase. The method, which can also be used for amplifying DNAs by non-rolling types of multiple displacement amplification, comprises incubating the reaction components in a small volume, e.g. about 10 μl or less, such as about 0.6 μl or less. The degree of amplification can be about 109 fold, or higher. A method for cell-free cloning of DNA, using the rolling circle amplification method of the invention, is described.

    摘要翻译： 本发明涉及例如通过等温滚圆机制扩增单链环状DNA分子（例如大小约5-6kb）的少量拷贝（例如单拷贝）的方法， 使用随机或部分随机引物和F29型DNA聚合酶。 也可以用于通过非滚动类型的多位置扩增来扩增DNA的方法包括将反应组分以小体积例如温育。 约10个或更少，例如约0.6毫摩尔或更少。 扩增程度可以是约109倍或更高。 描述了使用本发明的滚环扩增方法进行无细胞克隆DNA的方法。

    Amplification and cloning of single DNA molecules using rolling circle amplification
    13.
    发明授权
    Amplification and cloning of single DNA molecules using rolling circle amplification 有权
    使用滚环扩增扩增和克隆单个DNA分子

    公开(公告)号：US08497069B2

    公开(公告)日：2013-07-30

    申请号：US11919515

    申请日：2006-05-01

    申请人： Clyde A. Hutchison, III ,  Hamilton O. Smith

    发明人： Clyde A. Hutchison, III ,  Hamilton O. Smith

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12N15/1075 ,  C12N15/64 ,  C12Q1/6848 ,  C12Q2531/119 ,  C12Q2527/143 ,  C12Q2525/204 ,  C12Q2531/125

    摘要： The present invention relates, e.g., to a method for amplifying a small number of copies (e.g. a single copy) of a single-stranded circular DNA molecule (e.g. having a size of about 5-6 kb) by an isothermal rolling circle mechanism, using random or partially random primers and a F29-type DNA polymerase. The method, which can also be used for amplifying DNAs by non-rolling types of multiple displacement amplification, comprises incubating the reaction components in a small volume, e.g. about 10 μl or less, such as about 0.6 μl or less. The degree of amplification can be about 109 fold, or higher. A method for cell-free cloning of DNA, using the rolling circle amplification method of the invention, is described.

    摘要翻译： 本发明涉及例如通过等温滚圆机制扩增单链环状DNA分子（例如大小约5-6kb）的少量拷贝（例如单拷贝）的方法， 使用随机或部分随机引物和F29型DNA聚合酶。 也可以用于通过非滚动类型的多位置扩增来扩增DNA的方法包括将反应组分以小体积例如温育。 约10个或更少，例如约0.6毫摩尔或更少。 扩增程度可以是约109倍或更高。 描述了使用本发明的滚环扩增方法进行无细胞克隆DNA的方法。

    Installation of genomes or partial genomes into cells or cell-like systems
    14.
    发明授权
    Installation of genomes or partial genomes into cells or cell-like systems 有权
    将基因组或部分基因组安装到细胞或细胞样系统中

    公开(公告)号：US09434974B2

    公开(公告)日：2016-09-06

    申请号：US11644713

    申请日：2006-12-22

    申请人： John I. Glass ,  Lei Young ,  Carole Lartigue ,  Nacyra Assad-Garcia ,  Hamilton O. Smith ,  Clyde A. Hutchison ,  J. Craig Venter

    发明人： John I. Glass ,  Lei Young ,  Carole Lartigue ,  Nacyra Assad-Garcia ,  Hamilton O. Smith ,  Clyde A. Hutchison ,  J. Craig Venter

    IPC分类号： C12N15/74 ,  C12N15/63 ,  C12N15/10 ,  C12R1/35 ,  C12P21/02

    CPC分类号： C12P21/02 ,  C12N2510/00

    摘要： A method is provided for introducing a genome into a cell or cell-like system. The introduced genome may occur in nature, be manmade with or without automation, or may be a hybrid of naturally occurring and manmade materials. The genome is obtained outside of a cell with minimal damage. Materials such as a proteins, RNAs, polycations, nucleoid condensation proteins, or gene translation systems may accompany the genome. The genome is installed into a naturally occurring cell or into a manmade cell-like system. A cell-like system or synthetic cell resulting from the practice of the provided method may be designed and used to yield gene-expression products, such as desired proteins. By enabling the synthesis of cells or cell-like systems comprising a wide variety of genomes, accompanying materials and membrane types, the provided method makes possible a broader field of experimentation and bioengineering than has been available using prior art methods.

    摘要翻译： 提供了将基因组导入细胞或细胞样系统的方法。 引入的基因组可以在自然界中发生，在有或没有自动化的情况下是人造的，或者可以是天然存在的和人造的材料的混合物。 基因组以最小的损伤在细胞外获得。 材料如蛋白质，RNAs，聚阳离子，核仁缩合蛋白或基因翻译系统可能伴随着基因组。 将基因组安装到天然存在的细胞或人造细胞样系统中。 可以设计并使用由所提供的方法的实践产生的细胞样系统或合成细胞来产生基因表达产物，例如所需的蛋白质。 通过实现包含各种各样的基因组，伴随材料和膜类型的细胞或细胞样系统的合成，所提供的方法可能比使用现有技术方法可用的更广泛的实验和生物工程领域。

    METHODS FOR IN VITRO JOINING AND COMBINATORIAL ASSEMBLY OF NUCLEIC ACID MOLECULES
    18.
    发明申请
    METHODS FOR IN VITRO JOINING AND COMBINATORIAL ASSEMBLY OF NUCLEIC ACID MOLECULES 审中-公开
    核酸分子的体外联合和组合组装方法

    公开(公告)号：US20120053087A1

    公开(公告)日：2012-03-01

    申请号：US13035699

    申请日：2011-02-25

    申请人： Daniel G. Gibson ,  Hamilton O. Smith ,  Clyde A. Hutchison ,  Lei Young ,  J. Craig Venter

    发明人： Daniel G. Gibson ,  Hamilton O. Smith ,  Clyde A. Hutchison ,  Lei Young ,  J. Craig Venter

    IPC分类号： C12P19/34 ,  C40B40/06 ,  C07H21/04 ,  C12N9/12

    CPC分类号： C12P19/34 ,  C12N15/10 ,  C12N15/1027 ,  C12N15/64 ,  C12N15/66

    摘要： The present invention relates to methods of joining two or more double-stranded (ds) or single-stranded (ss) DNA molecules of interest in vitro, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule of each pair share a region of sequence identity. The method allows the joining of a large number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes. It can be used, e.g., to join synthetically produced sub-fragments of a gene or genome of interest. Kits for performing the method are also disclosed. The methods of joining DNA molecules may be used to generate combinatorial libraries useful to generate, for example, optimal protein expression through codon optimization, gene optimization, and pathway optimization.

    摘要翻译： 本发明涉及在体外连接两个或多个目的双链（ds）或单链（ss）DNA分子的方法，其中第一DNA分子的远端区域和第二DNA分子的近端区域 每对共享序列同一性区域。 该方法允许以预定顺序和取向连接大量DNA片段，而不使用限制酶。 其可以用于例如合成产生的感兴趣的基因或基因组的亚片段。 还公开了用于执行该方法的套件。 连接DNA分子的方法可用于产生组合文库，其可用于产生例如通过密码子优化，基因优化和途径优化的最佳蛋白质表达。

    Methods for in vitro joining and combinatorial assembly of nucleic acid molecules
    20.
    发明授权
    Methods for in vitro joining and combinatorial assembly of nucleic acid molecules 有权
    核酸分子的体外连接和组合装配方法

    公开(公告)号：US08968999B2

    公开(公告)日：2015-03-03

    申请号：US12371543

    申请日：2009-02-13

    申请人： Daniel G. Gibson ,  Hamilton O. Smith ,  Clyde A. Hutchison ,  Lei Young ,  J. Craig Venter

    发明人： Daniel G. Gibson ,  Hamilton O. Smith ,  Clyde A. Hutchison ,  Lei Young ,  J. Craig Venter

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/02 ,  C12N15/66 ,  C12N15/10 ,  C12N15/64

    CPC分类号： C12P19/34 ,  C12N15/10 ,  C12N15/1027 ,  C12N15/64 ,  C12N15/66

    摘要： The present invention relates to methods of joining two or more double-stranded (ds) or single-stranded (ss) DNA molecules of interest in vitro, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule of each pair share a region of sequence identity. The method allows the joining of a large number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes. It can be used, e.g., to join synthetically produced sub-fragments of a gene or genome of interest. Kits for performing the method are also disclosed. The methods of joining DNA molecules may be used to generate combinatorial libraries useful to generate, for example, optimal protein expression through codon optimization, gene optimization, and pathway optimization.

    摘要翻译： 本发明涉及在体外连接两个或多个目的双链（ds）或单链（ss）DNA分子的方法，其中第一DNA分子的远端区域和第二DNA分子的近端区域 每对共享序列同一性区域。 该方法允许以预定顺序和取向连接大量DNA片段，而不使用限制酶。 其可以用于例如合成产生的感兴趣的基因或基因组的亚片段。 还公开了用于执行该方法的套件。 连接DNA分子的方法可用于产生组合文库，其可用于产生例如通过密码子优化，基因优化和途径优化的最佳蛋白质表达。