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    Enhanced system for construction of adenovirus vectors
    2.
    发明授权
    Enhanced system for construction of adenovirus vectors 失效
    增强腺病毒载体构建体系

    公开(公告)号：US06756226B2

    公开(公告)日：2004-06-29

    申请号：US09981648

    申请日：2001-10-16

    Applicant: Frank L. Graham ,  Robin Parks ,  Philip Ng

    Inventor： Frank L. Graham ,  Robin Parks ,  Philip Ng

    IPC: C12N1564

    CPC classification number: C12N15/86 ,  A01K2217/05 ,  C12N2710/10343 ,  C12N2710/10351 ,  C12N2800/30

    Abstract: In the present invention, viruses, plasmids or both are constructed which contain viral DNA, at least one head-to-head ITR junction, and optionally, recombinase recognition sites positioned such that site-specific recombination between recombinase recognition sites in separate plasmids results in generation of infectious viral DNA at high-efficiency in cotransfected host cells that have been engineered to express a site-specific recombinase. Because of the high-efficiency and specificity of the Cre enzyme, suitably engineered plasmids can be readily recombined to produce infectious virus at high-efficiency in cotransfected 293 cells, without, at the same time, producing wild-type adenovirus, with the attendant problems for removal thereof. Use of recombinases besides Cre and recombinase recognition sites besides lox sites, and use of cells other than 293 cells are also disclosed and enabled, as are kits incorporating the site-specific vector system, as well as compositions and methods for using such compositions as vaccines or in gene therapeutic applications. Enhancements in the efficiency of both site-specific and homologous recombination are provided by inclusion of at least one head-to-head ITR junction.

    Abstract translation: 在本发明中，构建了包含病毒DNA，至少一个头对头ITR连接和任选的重组酶识别位点的病毒，质粒或二者，使得分离质粒中重组酶识别位点之间的位点特异性重组导致 在已经被工程化以表达位点特异性重组酶的共转染的宿主细胞中高效率地产生感染性病毒DNA。 由于Cre酶的高效率和特异性，合适的工程改造质粒可以在共转染293细胞中高效率地重组，产生感染性病毒，同时产生野生型腺病毒，伴随着问题 用于去除。 除了lox位点之外的Cre和重组酶识别位点以及使用除293细胞之外的细胞的重组酶的使用也被公开和启用，以及包含位点特异性载体系统的试剂盒以及使用这些组合物作为疫苗的组合物和方法 或在基因治疗应用中。 通过包含至少一个头对头ITR连接来提供位点特异性和同源重组的效率的增强。

    Method for transfer of DNA segments
    4.
    发明授权
    Method for transfer of DNA segments 失效
    DNA片段转移方法

    公开(公告)号：US06696278B1

    公开(公告)日：2004-02-24

    申请号：US09793372

    申请日：2001-02-26

    Applicant: Carsten-Peter Carstens

    Inventor： Carsten-Peter Carstens

    IPC: C12N1564

    CPC classification number: C12N15/66 ,  C12N15/10 ,  C12N15/64 ,  C12N15/70

    Abstract: The present invention provides a method of transfer of a gene of interest from a first vector to a product vector comprising contacting a first and second vector in vitro with a site-specific recombinase so as to generate a co-integrate vector comprising the components of the first and second vector, and introducing the co-integrate vector to a prokaryotic host cell so as to generate a product vector by rolling circle replication, comprising the gene of interest.

    Abstract translation: 本发明提供了将目的基因从第一载体转移到产物载体的方法，其包括使体外第一和第二载体与位点特异性重组酶接触，以便产生包含 第一和第二载体，并将共合并载体引入原核宿主细胞，以便通过包含感兴趣基因的滚环复制产生产物载体。

    Use of a virus DNA as promoter
    6.
    发明授权
    Use of a virus DNA as promoter 失效
    使用病毒DNA作为启动子

    公开(公告)号：US06303345B1

    公开(公告)日：2001-10-16

    申请号：US09462975

    申请日：2000-05-17

    Applicant: Wolfgang Rohde ,  Dieter Becker ,  John W. Randles ,  Alain Hehn ,  Francesco Salamini

    Inventor： Wolfgang Rohde ,  Dieter Becker ,  John W. Randles ,  Alain Hehn ,  Francesco Salamini

    IPC: C12N1564

    CPC classification number: C12N15/74 ,  C12N15/70 ,  C12N15/80 ,  C12N15/81 ,  C12N15/8216

    Abstract: Described is the characterization and the use of strong viral promoters for expressing genes, in bacteria and fungi, in particular yeasts. The invention is based on the surprising finding that CFDV DNA (coconut foliar decay virus DNA) and CFDV DNA fragments contain a region which is active as promoter even in bacteria and fungi, in particular yeasts, although they are derived from a virus which infects monocotyledonous plants. The activity of the promoters described in E. coli is distinctly higher than that of the CaMV 35S promoter, which is also active in bacteria.

    Abstract translation: 描述了强病毒启动子在细菌和真菌特别是酵母中表达基因的表征和用途。 本发明基于令人惊奇的发现，即CFDV DNA（椰子叶枯病病毒DNA）和CFDV DNA片段含有即使在细菌和真菌，特别是酵母中，作为启动子也是有活性的区域，尽管它们衍生自感染单子叶植物的病毒 植物。 在大肠杆菌中描述的启动子的活性明显高于CaMV 35S启动子的活性，其在细菌中也是活性的。

    High throughput DNA sequencing vector
    7.
    发明授权
    High throughput DNA sequencing vector 失效
    高通量DNA测序载体

    公开(公告)号：US06258571B1

    公开(公告)日：2001-07-10

    申请号：US09438142

    申请日：1999-11-10

    Applicant: Ilya Chumakov ,  Hiroaki Tanaka

    Inventor： Ilya Chumakov ,  Hiroaki Tanaka

    IPC: C12N1564

    CPC classification number: C12Q1/6841 ,  C12N9/16 ,  C12N9/2471 ,  C12N15/102 ,  C12N15/63 ,  C12N15/69 ,  C12Q1/6869 ,  C12Y302/01023

    Abstract: High throughput DNA sequencing vectors for generating nested deletions using enzymatic techniques and/or transposition-based techniques are disclosed. Methods of constructing contigs of long DNA sequences and methods of generating nested deletions are also disclosed. A truncated lacZ derivative useful in measuring the copy number of the lacZ derivative in a host cell is also disclosed.

    Abstract translation: 公开了用于使用酶技术和/或基于置换的技术产生嵌套缺失的高通量DNA测序载体。 还公开了构建长DNA序列的重叠群的方法和产生嵌套缺失的方法。 还公开了可用于测量宿主细胞中lacZ衍生物的拷贝数的截短的lacZ衍生物。

    Method for introducing a substance into a cell
    8.
    发明授权
    Method for introducing a substance into a cell 失效
    将物质引入细胞的方法

    公开(公告)号：US06562623B1

    公开(公告)日：2003-05-13

    申请号：US10031903

    申请日：2002-04-30

    Applicant: David Rickwood

    Inventor： David Rickwood

    IPC: C12N1564

    CPC classification number: C12N15/87 ,  C12M35/04

    Abstract: Provided is a method for introducing a substance into a cell, which method comprises: (a) generating one or more bubbles of a gas in a liquid medium comprising the cell, the bubbles being capable of forming a hole in the surface of the cell when one or more bubbles interact with the cell; and (b) introducing the substance into the cell.

    Abstract translation: 提供一种将物质引入电池的方法，该方法包括：（a）在包含电池的液体介质中产生一种或多种气体气泡，所述气泡能够在电池表面形成孔，当 一个或多个气泡与细胞相互作用; 和（b）将物质引入细胞中。

    Procedure for specific replacement of a copy of a gene present in the recipient genome by the integration of a gene different from that where the integration is made
    9.
    发明授权
    Procedure for specific replacement of a copy of a gene present in the recipient genome by the integration of a gene different from that where the integration is made 失效
    具体替换存在于受体基因组中的基因的拷贝通过整合不同于进行整合的基因的基因的程序

    公开(公告)号：US06528313B1

    公开(公告)日：2003-03-04

    申请号：US08301037

    申请日：1994-09-06

    Applicant: Hervé Le Mouellic ,  Philippe Brulet

    Inventor： Hervé Le Mouellic ,  Philippe Brulet

    IPC: C12N1564

    CPC classification number: A01K67/0275 ,  A01K2217/05 ,  A01K2227/105 ,  A61K38/00 ,  A61K48/00 ,  C07K14/47 ,  C12N15/85 ,  C12N15/8509 ,  C12N15/907 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2830/00 ,  C12N2830/85

    Abstract: The present invention relates to a process for targeted replacement of at least a part of an endogenous gene by at least a part of a foreign gene or targeted insertion of at least a part of a foreign gene at a targeted site in an endogenous gene in a cell by homologous recombination. This process includes (A) providing a vector which contains (1) at least a part of the foreign gene which is heterologous with respect to the endogenous gene; (2) a first flanking DNA sequence homologous to a first genomic sequence situated on one side of the part of the endogenous gene to be replaced or the targeted site; and (3) a second flanking DNA sequence homologous to a second genomic sequence situated on the other side of the part of the endogenous gene to be replaced or the targeted site, the foreign gene being located between the first and second flanking DNA sequences and is complementary to the part of the endogenous gene to be replaced; (B) transfecting a cell with the vector; (C) and selecting a transfected cell that contains the foreign gene at the targeted site or where the part of the endogenous gene is to be replaced.

    Abstract translation: 本发明涉及通过外源基因的至少一部分靶向置换至少一部分内源基因或至少部分外源基因靶向插入外源基因的内源基因的方法 细胞通过同源重组。 该方法包括（A）提供载体，其含有（1）外源基因的至少一部分相对于内源基因是异源的; （2）与位于待置换的内源基因的一部分的一侧上的第一基因组序列或靶位点同源的第一侧翼DNA序列; 和（3）与位于待替代的内源基因部分的另一侧的第二基因组序列同源的第二侧翼DNA序列或靶位点，外源基因位于第一和第二侧翼DNA序列之间，并且是 与待更换的内源基因的部分互补; （B）用载体转染细胞; （C），并且在靶位点选择含有外源基因的转染细胞，或者内源基因的一部分被置换的转染细胞。

    Nucleic acid sequence and methods for selectively expressing a protein in a target cell or tissue
    10.
    发明授权
    Nucleic acid sequence and methods for selectively expressing a protein in a target cell or tissue 有权
    核酸序列和用于在靶细胞或组织中选择性表达蛋白质的方法

    公开(公告)号：US06489141B1

    公开(公告)日：2002-12-03

    申请号：US09479645

    申请日：2000-01-07

    Applicant: Ian Hector Frazer ,  Jian Zhou

    Inventor： Ian Hector Frazer ,  Jian Zhou

    IPC: C12N1564

    CPC classification number: C12N7/00 ,  A61K2039/5258 ,  C07K14/005 ,  C07K14/43595 ,  C12N15/67 ,  C12N15/85 ,  C12N2710/20022 ,  C12N2710/20023 ,  C12N2710/20051 ,  C12N2830/00 ,  C12N2830/60 ,  C12Q1/6897

    Abstract: A synthetic polynucleotide and a method are disclosed for selectively expressing a protein in a target cell or tissue of a mammal. Selective expression is effected by replacing at least one existing codon of a parent polynucleotide encoding a protein of interest with a synonymous codon to produce a synthetic polynucleotide having altered translational kinetics compared to the parent polynucleotide. The synonymous codon is selected such that it has a higher translational efficiency in the target cell or tissue relative to one or more other cells or tissues of the mammal.

    Abstract translation: 公开了用于在哺乳动物的靶细胞或组织中选择性表达蛋白质的合成多核苷酸和方法。 通过使用同义密码子替换编码目标蛋白质的母体多核苷酸的至少一个现有密码子以产生与亲本多核苷酸相比具有改变的翻译动力学的合成多核苷酸来进行选择性表达。 选择同义密码子使得其相对于哺乳动物的一个或多个其它细胞或组织在靶细胞或组织中具有较高的翻译效率。

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