METHODS FOR IN VITRO JOINING AND COMBINATORIAL ASSEMBLY OF NUCLEIC ACID MOLECULES
    2.
    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

    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
    4.
    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

    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。

    Method of nucleic acid cassette assembly

    公开(公告)号:US10041060B2

    公开(公告)日:2018-08-07

    申请号:US11635355

    申请日:2006-12-06

    摘要: Methods are provided for constructing a synthetic genome, comprising generating and assembling nucleic acid cassettes comprising portions of the genome, wherein at least one of the nucleic acid cassettes is constructed from nucleic acid components that have been chemically synthesized, or from copies of the chemically synthesized nucleic acid components. In one embodiment, the entire synthetic genome is constructed from nucleic acid components that have been chemically synthesized, or from copies of the chemically synthesized nucleic acid components. Rational methods may be used to design the synthetic genome (e.g., to establish a minimal genome and/or to optimize the function of genes within a genome, such as by mutating or rearranging the order of the genes). Synthetic genomes of the invention may be introduced into vesicles (e.g., bacterial cells from which part or all of the resident genome has been removed, or synthetic vesicles) to generate synthetic cells. Synthetic genomes or synthetic cells may be used for a variety of purposes, including the generation of synthetic fuels, such as hydrogen or ethanol.

    Installation of genomes or partial genomes into cells or cell-like systems
    10.
    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

    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。