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71. 发明申请

WO2007047859A3 RATIONALLY-DESIGNED MEGANUCLEASES WITH ALTERED SEQUENCE SPECIFICITY AND DNA-BINDING AFFINITY 审中-公开
标题翻译：具有改变的序列特异性和DNA结合亲和力的方案设计的重组
公开(公告)号：WO2007047859A3
公开(公告)日：2007-11-15
申请号：PCT/US2006040919
申请日：2006-10-18
申请人： PREC BIOSCIENCES , HELLINGA HOMME W , SMITH JAMES JEFFERSON , JANTZ DEREK
发明人： HELLINGA HOMME W , SMITH JAMES JEFFERSON , JANTZ DEREK
IPC分类号： C12N9/22 , C12N15/90
CPC分类号： C12N9/22 , A61K38/465 , A61K48/00 , C12N15/8213 , C12N15/902 , C12N15/905 , C12N15/907
摘要： Rationally-designed LAGLIDADG meganucleases and methods of making such meganucleases are provided. In addition, methods are provided for using the meganucleases to generate recombinant cells and organisms having a desired DNA sequence inserted into a limited number of loci within the genome, as well as methods of gene therapy, for treatment of pathogenic infections, and for in vitro applications in diagnostics and research.
摘要翻译：提供合理设计的LAGLIDADG大范围核酸酶和制备这种大范围核酸酶的方法。此外，提供了使用大范围核酸酶产生重组细胞和生物体的方法，所述重组细胞和生物体具有插入基因组内有限数量基因座的所需DNA序列，以及基因治疗方法，用于治疗病原体感染和体外应用于诊断和研究。

72. 发明申请

WO2007115886A1 IMPROVED METHOD FOR HOMOLOGOUS RECOMBINATION IN EUKARYOTIC CELLS 审中-公开
标题翻译：改造方法用于真核细胞中的同源重组
公开(公告)号：WO2007115886A1
公开(公告)日：2007-10-18
申请号：PCT/EP2007/052392
申请日：2007-03-14
申请人： DSM IP ASSETS B.V. , BERG, VAN DEN, Marco Alexander , KERKMAN, Richard , TOUW-RIEL, Hesselien
发明人： BERG, VAN DEN, Marco Alexander , KERKMAN, Richard , TOUW-RIEL, Hesselien
IPC分类号： C12N15/80 , C12N15/90
CPC分类号： C12N15/815 , C12N15/905 , C12N2800/30 , C12N2800/70
摘要： The present invention discloses a method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a desired replacement sequence, comprising: modifying a parent eukaryotic cell with a preference for NHR to provide a eukaryotic cell having an increased HR/NHR ratio as compared to the parent cell, providing a DNA molecule comprising a first DNA fragment comprising a desired replacement sequence flanked at its 5' and 3' sides by DNA sequences substantially homologous to sequences of the chromosomal DNA flanking the target sequence and a second DNA fragment comprising an expression cassette comprising a gene encoding a selection marker and regulatory sequences functional in the eukaryotic cell operably linked thereto; transforming the modified eukaryotic cells with the DNA molecule; growing the cells to obtain transformed progeny cells having the DNA molecule inserted into the chromosome, deselecting the progeny cells in which the DNA molecule is inserted in the chromosome via a non-homologous recombination event by expression of the selection marker; and obtaining cells wherein the target sequence in the chromosomal DNA sequence is replaced by the desired replacement sequence.
摘要翻译：本发明公开了一种构建真核细胞的方法，所述真核细胞在染色体DNA序列中以期望的置换序列替代具有靶序列，其包括：以优选NHR修饰亲本真核细胞以提供具有增加的HR / NHR比的真核细胞与亲本细胞相比，提供DNA分子，其包含第一DNA片段，所述第一DNA片段包含在其5'和3'侧侧翼的所需替换序列，所述DNA序列与靶序列侧翼的染色体DNA的序列基本同源的DNA序列和第二DNA片段包括表达盒，其包含编码选择标记的基因和在可操作地连接到其上的在真核细胞中功能的调节序列; 用DNA分子转化修饰的真核细胞; 生长细胞以获得具有插入到染色体中的DNA分子的转化后代细胞，通过选择标记的表达，通过非同源重组事件去除选择DNA分子插入到染色体中的子代细胞; 并获得细胞，其中染色体DNA序列中的靶序列被所需的置换序列替代。

73. 发明申请

WO2007063919A1 染色体改変方法审中-公开
标题翻译：修饰色素的方法
公开(公告)号：WO2007063919A1
公开(公告)日：2007-06-07
申请号：PCT/JP2006/323852
申请日：2006-11-29
申请人：旭硝子株式会社 , 平島匡太郎 , 東田英毅 , 浜祐子
发明人：平島匡太郎 , 東田英毅 , 浜祐子
IPC分类号： C12N15/00 , C12N15/09 , C12Q1/68
CPC分类号： C12N15/815 , C12N15/1079 , C12N15/1082 , C12N15/905
摘要： It is intended to provide a widely usable method of modifying chromosome without resorting to any specific enzyme or sequence and a method of identifying a gene that is essentially required in cell proliferation. First, a selection marker gene (B) allowing negative selection is integrated into the upstream or downstream of a target DNA region (X) and then the selection marker gene (B) and the target DNA region (X) are sandwiched between duplicated sequences. It is preferable that a DNA fragment having a sequence (A') (which is a sequence being substantially the same as a specific sequence (A) located in the downstream of the target DNA region) and the selection marker gene (B) as described above is constructed and then this DNA fragment is integrated into the upstream of the target DNA region (X). Next, homologous recombination and negative selection are conducted so that the target DNA region (X) can be easily removed from the chromosome. In the case where the chromosome contains no target DNA region (X) after the above-described treatment, it can be understood that the target DNA region (X) contains no gene that is essentially required in cell proliferation under the culture condition.
摘要翻译：本发明旨在提供一种广泛可用的修饰染色体的方法，而不涉及任何特定的酶或序列，以及鉴定细胞增殖中基本上需要的基因的方法。首先，将允许阴性选择的选择标记基因（B）整合到靶DNA区域（X）的上游或下游，然后将选择标记基因（B）和靶DNA区域（X）夹在复制序列之间。优选具有序列（A'）（其位于靶DNA区域下游的与特定序列（A）基本相同的序列）的DNA片段和所述选择标记基因（B）构建上述，然后将该DNA片段整合到靶DNA区域（X）的上游。接下来，进行同源重组和阴性选择，使得靶DNA区域（X）可以容易地从染色体上去除。在染色体在上述处理后不含目标DNA区域（X）的情况下，可以理解，目标DNA区域（X）不含有在培养条件下细胞增殖基本上必需的基因。

74. 发明申请

WO02052026A8 NUCLEIC ACID INTEGRATION IN EUKARYOTES 审中-公开
标题翻译：核酸的核酸整合
公开(公告)号：WO02052026A8
公开(公告)日：2003-11-27
申请号：PCT/NL0100936
申请日：2001-12-21
申请人： UNIV LEIDEN , STICHTING BINAIR VECTOR SYSTEE , HOOYKAAS PAUL JAN JACOB , VAN ATTIKUM HAICO , BUNDOCK PAUL
发明人： HOOYKAAS PAUL JAN JACOB , VAN ATTIKUM HAICO , BUNDOCK PAUL
IPC分类号： C12N15/81 , C12N15/82 , C12N15/90
CPC分类号： C12N15/8213 , C12N15/81 , C12N15/902 , C12N15/905
摘要： The invention relate to the field of molecular biology and cell biology. It particularly relates to methods to direct integration of a nucleic acid of interest towards homologous recombination and uses thereof. The present invention discloses factors involved in integration of a nucleic acid by illegitimate recombination which provides a method to direct integration of a nucleic acid of interest to a pre-determined site, whereby said nucleic acid has homology at or around the said pre-determined site, in a eukaryote with a preference for non-homologous recombination comprising steering an integration pathway towards homologous recombination. Furthermore, the invention provides a method to direct integration of a nucleic acid of interest to a sub-telomeric and/or telomeric region in a eukaryote with a preference for non-homologous recombination.
摘要翻译：本发明涉及分子生物学和细胞生物学领域。它特别涉及将感兴趣的核酸直接整合到同源重组及其用途的方法。本发明公开了涉及通过非法重组整合核酸的因素，其提供了将感兴趣的核酸直接整合到预定位点的方法，由此所述核酸在所述预定位点处或周围具有同源性，在优先选择非同源重组的真核生物中，包括转向同源重组的整合途径。此外，本发明提供了一种直接将目的核酸整合到真核生物中的亚端粒和/或端粒区域并优先选择非同源重组的方法。

75. 发明申请

WO0222811A3 MODULATION OF MEIOTIC RECOMBINATION 审中-公开
标题翻译：调节重建
公开(公告)号：WO0222811A3
公开(公告)日：2003-10-02
申请号：PCT/CA0101306
申请日：2001-09-12
申请人： CA MINISTER AGRICULTURE & FOOD , ROZWADOWSKI KEVIN L , LYDIATE DEREK J
发明人： ROZWADOWSKI KEVIN L , LYDIATE DEREK J
IPC分类号： C12N15/82 , C12N15/873 , C12N15/90 , C12N15/01 , A01H1/02 , A01K67/02 , A01K67/027 , C07K14/395 , C12N3/00 , C12Q1/68
CPC分类号： C12N15/81 , C12N15/8213 , C12N15/873 , C12N15/905 , C12N15/907
摘要： The invention provides methods of modifying the level of expression or functional activity of factors such as enzymes or other catalytic proteins or structural proteins, alone or in concert, to modify the frequency of meiotic homologous recombination involving the exchange of genetic information between non-sister chromatids from homologous maternal and paternal chromosomes. The steps at which modulation may occur include: homologous chromosome pairing, double-strand break formation; resection; strand invasion; branch migration; and resolution. Methods of plant and animal breeding are also provided that utilize the modulation of meiotic homologous recombination.
摘要翻译：本发明提供了单独或一致地修饰诸如酶或其他催化蛋白或结构蛋白等因素的表达水平或功能活性的方法，以修饰涉及非姐妹染色单体之间的遗传信息交换的减数分裂同源重组的频率来自同源的母体和父系染色体。可能发生调节的步骤包括：同源染色体配对，双链断裂形成; 切除; 线入侵; 分支迁移; 和决议。还提供了利用减数分裂同源重组调控的植物和动物育种方法。

76. 发明申请

WO02052026A3 NUCLEIC ACID INTEGRATION IN EUKARYOTES 审中-公开
标题翻译：核酸的核酸整合
公开(公告)号：WO02052026A3
公开(公告)日：2002-09-06
申请号：PCT/NL0100936
申请日：2001-12-21
申请人： UNIV LEIDEN , STICHTING BINAIR VECTOR SYSTEE , HOOYKAAS PAUL JAN JACOB , VAN ATTIKUM HAICO , BUNDOCK PAUL
发明人： HOOYKAAS PAUL JAN JACOB , VAN ATTIKUM HAICO , BUNDOCK PAUL
IPC分类号： C12N15/81 , C12N15/82 , C12N15/90
CPC分类号： C12N15/8213 , C12N15/81 , C12N15/902 , C12N15/905
摘要： The invention relate to the field of molecular biology and cell biology. It particularly relates to methods to direct integration of a nucleic acid of interest towards homologous recombination and uses thereof. The present invention discloses factors involved in integration of a nucleic acid by illegitimate recombination which provides a method to direct integration of a nucleic acid of interest to a pre-determined site, whereby said nucleic acid has homology at or around the said pre-determined site, in a eukaryote with a preference for non-homologous recombination comprising steering an integration pathway towards homologous recombination. Furthermore, the invention provides a method to direct integration of a nucleic acid of interest to a sub-telomeric and/or telomeric region in a eukaryote with a preference for non-homologous recombination.
摘要翻译：本发明涉及分子生物学和细胞生物学领域。它特别涉及将感兴趣的核酸直接整合到同源重组及其用途的方法。本发明公开了涉及通过非法重组整合核酸的因素，其提供了将感兴趣的核酸直接整合到预定位点的方法，由此所述核酸在所述预定位点处或周围具有同源性，在优先选择非同源重组的真核生物中，包括转向同源重组的整合途径。此外，本发明提供了一种直接将目的核酸整合到真核生物中的亚端粒和/或端粒区域并优先选择非同源重组的方法。

77. 发明申请

WO01051646A2 METHODS FOR PRODUCING A POLYPEPTIDE USING A CRIPPLED TRANSLATIONAL INITIATOR SEQUENCE 审中-公开
标题翻译：使用切片翻译序列产生多肽的方法
公开(公告)号：WO01051646A2
公开(公告)日：2001-07-19
申请号：PCT/US2001/001102
申请日：2001-01-12
IPC分类号： C12N15/80 , C12N15/90 , C12N15/67 , C12N15/52 , C12N15/65
CPC分类号： C12N15/905 , C12N15/80
摘要： The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a fungal host cell in a medium conducive for the production of the polypeptide; and (b) isolating the polypeptide from the cultivation medium; wherein the fungal host cell comprises a first nucleic acid sequence encoding the polypeptide in tandem with a second nucleic acid sequence comprising a crippled translational initiator sequence operably linked to a gene encoding a selectable marker, wherein the copy number of the first nucleic acid sequence has been increased by culturing the cell under conditions that select for multiple copies of the selectable marker. The present invention also relates to such fungal host cells and methods for obtaining such fungal host cells. The present invention further relates to nucleic acid constructs and vectors comprising a crippled translational initiator sequence operably linked to a gene encoding a selectable marker.
摘要翻译：本发明涉及产生多肽的方法，包括：（a）在有助于产生多肽的培养基中培养真菌宿主细胞; 和（b）从培养基中分离多肽; 其中所述真菌宿主细胞包含编码所述多肽的第一核酸序列与第二核酸序列串联，所述第二核酸序列包含与编码可选择标记的基因可操作地连接的残基翻译起始序列，其中所述第一核酸序列的拷贝数已经被通过在选择可选择标记的多个拷贝的条件下培养细胞而增加。本发明还涉及这种真菌宿主细胞和获得这种真菌宿主细胞的方法。本发明还涉及核酸构建体和载体，其包含与编码可选择标记的基因可操作地连接的残基翻译起始序列。

78. 发明申请

WO00053781A1 GENERATION OF CONDITIONAL YEAST MUTANTS, METHODS AND REAGENTS RELATED THERETO 审中-公开
标题翻译：条件YEAST MUTANTS，相关方法和试剂的产生
公开(公告)号：WO00053781A1
公开(公告)日：2000-09-14
申请号：PCT/US2000/006514
申请日：2000-03-13
IPC分类号： C12N1/19 , C12N15/31 , C12N15/81 , C12N15/90 , C12Q1/00
CPC分类号： C12N15/815 , C12N15/905
摘要： The invention features a method for systematically generating mutant strains of obligate haploid and obligate diploid yeast, having condition-sensitive lethal mutations of a gene which is otherwise essential to cell viability under all conditions of cell growth. Conditional mutants make possible the analysis of physiological changes caused by inactivation of a gene or a gene product and can be used to address the function of any gene. In an exemplary embodiment, the method of the present invention can be generally characterized as follows: (i) providing a Candida strain whichis heterozygous for a loss-of-function mutation of an allele of an essential gene; and (ii) introducing a conditionally sensitive homolog of the gene, e.g., with conditional-sensitive protein or conditional-sensitive transcriptional regulatory sequence, at the functional allele by integrative transformation.
摘要翻译：本发明的特征在于一种用于系统地产生专性单倍体和专性二倍体酵母的突变株的方法，其具有在细胞生长的所有条件下对细胞活力至关重要的基因的条件敏感致死突变。有条件的突变体使得能够分析由基因或基因产物失活引起的生理变化，并可用于解决任何基因的功能。在一个示例性实施方案中，本发明的方法的一般特征如下：（i）提供对必需基因的等位基因的功能丧失突变杂合的假丝酵母属菌株; 和（ii）通过整合转化在功能等位基因上引入基因的条件敏感同源物，例如条件敏感蛋白或条件敏感性转录调控序列。

79. 发明申请

WO9629413A2 DNA ENCODING HUMAN PAPILLOMA VIRUS TYPE 18 审中-公开
标题翻译： DNA编码人类斑马鱼病毒18型
公开(公告)号：WO9629413A2
公开(公告)日：1996-09-26
申请号：PCT/US9603649
申请日：1996-03-18
申请人： MERCK & CO INC , HOFMANN KATHRYN J , JANSEN KATHRIN U , NEEPER MICHAEL P , JOYCE JOSEPH G , GEORGE HUGH A
发明人： HOFMANN KATHRYN J , JANSEN KATHRIN U , NEEPER MICHAEL P , JOYCE JOSEPH G , GEORGE HUGH A
IPC分类号： C12N15/09 , A61K39/00 , A61K39/12 , A61K48/00 , A61P31/12 , C07K14/025 , C07K16/08 , C12N7/00 , C12N7/04 , C12N9/10 , C12N9/60 , C12N15/33 , C12N15/37 , C12N15/81 , C12N15/90 , C12P21/02 , C12R1/865 , C12R1/91
CPC分类号： C07K14/005 , A61K39/00 , C07K2319/00 , C12N7/00 , C12N9/1051 , C12N9/60 , C12N15/905 , C12N2710/20022 , C12N2710/20023
摘要： The present invention is directed to DNA molecules encoding purified human papillomavirus type 18 and derivatives thereof.
摘要翻译：本发明涉及编码18型纯化人乳头状瘤病毒的DNA分子及其衍生物。

80. 发明申请

WO1992001069A1 LIBRARY SCREENING METHOD 审中-公开
标题翻译：图书馆筛选方法
公开(公告)号：WO1992001069A1
公开(公告)日：1992-01-23
申请号：PCT/US1991004926
申请日：1991-07-12
申请人： TRANSKARYOTIC THERAPIES, INC.
发明人： TRANSKARYOTIC THERAPIES, INC. , TRECO, Douglas, A. , MILLER, Allan, M.
IPC分类号： C12Q01/68
CPC分类号： C12N15/1082 , C12N15/81 , C12N15/905 , C12Q1/02 , C12Q1/68
摘要： Materials and methods for homologous-recombination screening of DNA libraries constructed in a eukaryotic host and methods for homologous-recombination chromosome walking for isolating overlapping DNA sequences for building an extended physical map of a chromosomal region.
摘要翻译：用于在真核宿主中构建的DNA文库的同源重组筛选的材料和方法以及用于分离重叠DNA序列以构建染色体区域的扩展物理图的同源重组染色体步行方法。

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