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公开(公告)号：US07838287B2
公开(公告)日：2010-11-23
申请号：US10466960
申请日：2002-01-25
CPC分类号： C12N15/1034 , C12N15/1086 , C12N15/66
摘要： The present invention relates to combinatorial gene expression libraries and methods for making these. Such libraries are useful in discovery of novel and/or enhanced metabolic pathways leading to the production of novel compounds for e.g drug discovery and/or to the production of known compounds in novel quantities or in novel compartments of the cells. The expression libraries in particular are composed of host cells capable of co-ordinated and controllable expression of large numbers of heterologous genes in the host cells.
摘要翻译： 本发明涉及组合基因表达文库及其制备方法。 这样的文库可用于发现新的和/或增强的代谢途径，导致生成用于例如药物发现的新化合物和/或以新的量或新颖的细胞室中生产已知化合物。 表达文库特别由能够在宿主细胞中协调和可控制表达大量异源基因的宿主细胞组成。
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公开(公告)号：US20060252156A1
公开(公告)日：2006-11-09
申请号：US10523007
申请日：2003-08-01
CPC分类号： C12N15/1093 , C12N15/02 , C12N15/1027
摘要： The present invention relates to methods of mixing large numbers of heterologous genes, which are located on artificial chromsomes. The methods of the present invention are useful for evolution of cells and whole genomes to acquire new functionality(ies), such as the ability to synthesise novel secondary metabolites and/or the evolution of novel metabolic pathways. In particular the methods may involve mating or fusing two populations of cells, each population comprising different combinations of heterologous genes. Subsequently, desirable cells may be selected. In embodiments of the invention the heterologous genes are comprised within expression cassettes. The expression cassette can be in the form of a concatermer, and can be cut out using a restriction enzyme in order to make new combinations of expression cassettes.
摘要翻译： 本发明涉及混合大量位于人造染色体上的异源基因的方法。 本发明的方法可用于细胞和整个基因组的进化以获得新的功能性，例如合成新的次级代谢物的能力和/或新型代谢途径的进化。 特别地，所述方法可涉及交配或融合两个细胞群，每个群体包含异源基因的不同组合。 随后，可以选择期望的细胞。 在本发明的实施方案中，异源基因包含在表达盒内。 表达盒可以是连接体的形式，并且可以使用限制性内切酶切割以形成表达盒的新组合。
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3.发明申请Methods for multiple parameters screening and evolution of cells to produce small molecules with multiple functionalities 审中-公开用于多参数筛选和细胞进化的方法来产生具有多种功能的小分子公开(公告)号：US20050164162A1
公开(公告)日：2005-07-28
申请号：US10502489
申请日：2003-01-24
CPC分类号： C12Q1/18 , C12N15/1034 , C12N15/1086 , G01N33/5008 , G01N33/5011 , G01N33/5014 , G01N33/502 , G01N33/5097
摘要： The present invention relates to the field of selecting cells producing compounds that fulfil at least 2 predetermined functionalities. The invention primarily focuses on methods for screening of cells for two or more functionalities related to small molecules generated by host cells with novel gene assemblies. The methods in one aspect of the invention are combined with methods for evolution of cells to acquire at least two predetermined functionalities related to small molecules. Furthermore, the invention relates to methods for generation of lead compounds that are selected for a primary function and at least one further secondary function. The methods according to the invention may in particular be used in high troughput screening.
摘要翻译： 本发明涉及选择产生满足至少2个预定功能的化合物的细胞的领域。 本发明主要关注用于筛选与由具有新基因组件的宿主细胞产生的小分子相关的两个或多个功能的细胞的方法。 将本发明的一个方面中的方法与细胞进化的方法结合以获得与小分子相关的至少两个预定的功能。 此外，本发明涉及用于产生主要功能和至少一种其它次要功能的铅化合物的生产方法。 根据本发明的方法可以特别用于高水平筛选。
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公开(公告)号：US08008459B2
公开(公告)日：2011-08-30
申请号：US10466959
申请日：2002-01-25
CPC分类号： C12N15/85 , C12N15/66 , C12N15/70 , C12N15/81 , C12N2800/20 , C12N2800/206
摘要： In the present invention are disclosed concatemers of concatenated expression cassettes and vectors that enable the synthesis of such concatemers. The concatemer comprises in the 5′→3′ direction a cassette of nucleotide sequence of the general formula [rs2-SP-PR-X-TR-SP-rs1]n wherein rs1 and rs2 together denote a functional restriction site, SP individually denotes a spacer of at least two nucleotide bases, PR denotes a promoter, capable of functioning in a cell, X denotes an expressible nucleotide sequence, TR denotes a terminator, and SP individually denotes a spacer of at least two nucleotide bases, and n>/=2, and wherein at least a first cassette is different from a second cassette. The main purpose of these concatemers is the controllable and co-ordinated expression of large numbers of heterologous genes in a single host. Furthermore, the invention relates to a concatemer of cassettes of nucleotide sequences and a method for preparing the concatemers. In a further aspect, the invention relates to transgenic host cells comprising at least one concatemer according to the invention, as well as to a method for preparing the transgenic host cells. Finally, the invention relates to a vector comprising a cassette of nucleotides, a method for preparing said vector, a nucleotide library comprising at least two primary vectors each comprising a cassette of nucleotides, a method for preparing the library.
摘要翻译： 在本发明中公开了连接的表达盒和能够合成这种并发物的载体的并列物质。 连接体在5'→3'方向上包含通式[rs2-SP-PR-X-TR-SP-rs1] n的核苷酸序列的盒，其中rs1和rs2一起表示功能性限制性位点，SP分别表示 至少两个核苷酸碱基的间隔区，PR表示能够在细胞中起作用的启动子，X表示可表达的核苷酸序列，TR表示终止子，SP表示至少两个核苷酸碱基的间隔区，n> = 2，并且其中至少第一盒不同于第二盒。 这些结合物的主要目的是在单个宿主中可控和协调的大量异源基因的表达。 此外，本发明涉及核苷酸序列的盒的连接体和制备该并列物质的方法。 在另一方面，本发明涉及包含根据本发明的至少一种连接体的转基因宿主细胞，以及制备转基因宿主细胞的方法。 最后，本发明涉及包含核苷酸盒的载体，制备所述载体的方法，包含至少两个主要载体的核苷酸文库，每个载体包含核苷酸盒，制备文库的方法。
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公开(公告)号：US20120094870A1
公开(公告)日：2012-04-19
申请号：US12902587
申请日：2010-10-12
CPC分类号： C12N15/1034 , C12N15/1086 , C12N15/66
摘要： The present invention relates to combinatorial gene expression libraries and methods for making these. Such libraries are useful in discovery of novel and/or enhanced metabolic pathways leading to the production of novel compounds for e.g., drug discovery and/or to the prosecution of known compounds in novel quantities or in novel compartments of the cells. The expression libraries in particular are composed of host cells capable of co-ordinated and controllable expression of large numbers of heterologous genes in the host cells.
摘要翻译： 本发明涉及组合基因表达文库及其制备方法。 这样的文库可用于发现新的和/或增强的代谢途径，导致新的化合物的生产，例如药物发现和/或检测新颖数量的或新颖的细胞室中的已知化合物。 表达文库特别由能够在宿主细胞中协调和可控制表达大量异源基因的宿主细胞组成。
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6.发明申请公开(公告)号：US20050158860A1
公开(公告)日：2005-07-21
申请号：US10470062
申请日：2002-01-25
IPC分类号： C12N15/09 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12N15/10 , C12P21/02 , C12R1/66 , C12R1/865 , C12N15/74 , C12N1/18
CPC分类号： C12N15/1027 , C12N15/1051 , C12N15/1079 , C12N15/1086
摘要： The present invention relates to evolution of a cell or a composition of cells having a desired property or functionally. The principle behind the evolution of cells according to the invention is to produce a great diversity of genes in each cell subjected to evolution and a great diversity of genes among the cells in a composition according to the invention and to exchange the genes between the cells from time to time. In preferred embodiments the genes are arranged in expression cassettes in concatemers in the cells, as well as in artificial chromosomes. Said methods comprising the steps of a) obtaining a composition of cells, at least one cell of said composition comprising a1) at least two expressible nucleotide sequences, at least one of said sequences being incorporated into an artificial chromosome in the cell and/or a2) at least two expression cassettes of the following formula: [rs2-SP-PR-X-TR-SP-rs11 wherein rs1 and rs2 together denotes a restriction site, SP individually denotes a spacer, PR denotes a promoter, capable of functionning in the first cell, X denotes an expressible nucleotide sequence, TR denotes a terminator, and/or a3) at least two expressible nucleotide sequences, said sequences being heterologous to the cell, determining at least one screening functionality. The genes are preferably co-ordinately controllable for increasing diversity. The desired property of functionality is preferably a compound, or a series of compounds synergistically acting to each other.
摘要翻译： 本发明涉及具有所需性质或功能的细胞的细胞或组成的进化。 根据本发明的细胞进化背后的原理是在根据本发明的组合物中经历进化和每个细胞中基因的多样性的每个细胞中产生大量的基因，并且将细胞之间的基因交换 时不时 在优选的实施方案中，将基因排列在细胞中以及人造染色体中的并列的表达盒中。 所述方法包括以下步骤：a）获得细胞组合物，所述组合物的至少一个细胞包含a1）至少两个可表达的核苷酸序列，所述序列中的至少一个被并入细胞和/或a2中的人造染色体 ）至少两个下式的表达盒：[rs2-SP-PR-X-TR-SP-rs11其中rs1和rs2一起表示限制性位点，SP分别表示间隔基，PR表示能够在 第一个细胞，X表示可表达的核苷酸序列，TR表示终止子，和/或a3）至少两个可表达的核苷酸序列，所述序列与细胞异源，确定至少一个筛选功能。 这些基因优选可协同控制以增加多样性。 功能性所需的性质优选是化合物或一系列协同作用的化合物。
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