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1. 发明申请

US20100173364A1 Repair of Nucleic Acids for Improved Amplification 审中-公开
标题翻译：用于改进扩增的核酸修复
公开(公告)号：US20100173364A1
公开(公告)日：2010-07-08
申请号：US12296827
申请日：2007-04-11
申请人： Thomas C. Evans, JR. , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan , Rebecca Kucera
发明人： Thomas C. Evans, JR. , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan , Rebecca Kucera
IPC分类号： C12P19/34 , C12N9/00 , C12N9/10
CPC分类号： C12Q1/686 , C12Q2521/514 , C12Q2521/501 , C12Q2521/301 , C12Q2525/119
摘要： Methods and compositions are provided for repairing a polynucleotide so that it can be copied with improved fidelity and/o yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a DNA ligase and an effective amount of at least one endonuclease as well as a cofactor selected from NAD+ or ATP.
摘要翻译：提供了用于修复多核苷酸的方法和组合物，使得其可以在例如扩增反应中以改进的保真度和/或产率复制。这包括使用包括DNA连接酶和有效量的至少一种内切核酸酶以及选自NAD +或ATP的辅因子的反应混合物。

2. 发明授权

US08158388B2 Repair of nucleic acids for improved amplification 有权
标题翻译：修复核酸以改善扩增
公开(公告)号：US08158388B2
公开(公告)日：2012-04-17
申请号：US11401826
申请日：2006-04-11
申请人： Thomas C. Evans , Lixin Chen , Chudi Guan , Rebecca Kucera , Barton Slatko , Romualdas Vaisvila
发明人： Thomas C. Evans , Lixin Chen , Chudi Guan , Rebecca Kucera , Barton Slatko , Romualdas Vaisvila
IPC分类号： C12P19/34
CPC分类号： C12Q1/6844 , C12P19/34 , C12Q1/6848 , C12Q1/686 , C12Q2521/101 , C12Q2521/501 , C12Q2521/331 , C12Q2521/301 , C12Q2521/514
摘要： Methods and compositions are provided for repairing a polynucleotide so that it can be copied with improved fidelity and/or yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI.The reaction mixture may further contain an AP endonuclease and a polymerase. If used, these enzymes may be selected according to their ability to withstand high temperatures. For example, the reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture in as much as the repair is effected rapidly and prolonged incubation is not generally adverse.
摘要翻译：提供了用于修复多核苷酸的方法和组合物，使得其可以在例如扩增反应中以改进的保真度和/或产率复制。这包括使用包含连接酶和选自NAD +或ATP的辅因子并在不存在核酸内切酶VI的情况下将多核苷酸与反应混合物一起温育的反应混合物。反应混合物还可以含有AP内切核酸酶和聚合酶。如果使用，这些酶可以根据其耐受高温的能力来选择。例如，反应混合物可以在多核苷酸合成反应之前使用，在这种情况下可以使用不是嗜热的酶。维修反应对于酶混合物中的几秒钟，几分钟或几小时的孵育时间并不敏感，只要修复迅速进行并且延长的孵育通常不是不利的。

3. 发明授权

US06358724B1 Isolation and composition of novel glycosidases 失效
标题翻译：新型糖苷酶的分离和组成
公开(公告)号：US06358724B1
公开(公告)日：2002-03-19
申请号：US09883800
申请日：2001-06-18
申请人： Sharon T. Wong-Madden , Ellen P. Guthrie , David Landry , Christopher H. Taron , Chudi Guan , Phillips W. Robbins
发明人： Sharon T. Wong-Madden , Ellen P. Guthrie , David Landry , Christopher H. Taron , Chudi Guan , Phillips W. Robbins
IPC分类号： C12N938
CPC分类号： C12Y302/0105 , C12N9/2471 , C12Y302/01023
摘要： Substantially pure glycosidases capable for cleaving selected glycosidic bonds have been described including glycosidases isolated from Xanthomonas and recombinant glycosidases. Substrate specificity of isolated enzymes have been identified for GlcNac&bgr;1-X, Gal&agr;1-3R, Gal&agr;1-6R, Gal&bgr;1-3R, Fuc&agr;-2R, Fuc&agr;1-3R, Fuc&agr;1-4R, Man&agr;1-2R, Man&agr;1-3R, Man&agr;1-6R, Man&bgr;1-4R, Xyl&bgr;1-2R, Glc&bgr;1-4R, and Gal&bgr;1-4R providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.
摘要翻译：已经描述了能够切割选择的糖苷键的基本上纯的糖苷酶，其包括从黄单胞菌属和重组糖苷酶分离的糖苷酶。已经鉴定了GlcNacbeta1-X，Galalpha1-3R，Galalpha1-6R，Galbeta1-3R，Fucalpha-2R，Fucalpha1-3R，Fucalpha1-4R，Manalpha1-2R，Manalpha1-3R，Manalpha1-6R，Manbeta1的分离的酶的底物特异性 -4R，Xylbeta1-2R，Glcbeta1-4R和Galbeta1-4R，其提供了选择性地切割碳水化合物底物中的糖苷键并形成修饰的碳水化合物的改善的能力。

4. 发明授权

US07851192B2 Modified DNA cleavage enzymes and methods for use 有权
标题翻译：修饰的DNA切割酶和使用方法
公开(公告)号：US07851192B2
公开(公告)日：2010-12-14
申请号：US10585964
申请日：2004-11-22
申请人： Chudi Guan , Sanjay Kumar , Rebecca Kucera
发明人： Chudi Guan , Sanjay Kumar , Rebecca Kucera
IPC分类号： C12N9/16 , C12Q1/44 , C12Q1/68 , C07K14/00 , C12N15/00 , C12N1/21 , C12P21/00 , C07H21/00
CPC分类号： C12N9/22
摘要： Compositions and methods are provided that relate to a modified DNA cleaving enzyme having at least 35% amino acid sequence identity with T7 Endo I. The modified enzyme includes two catalytic centers separated by a β-bridge where the β-bridge contains at least one mutation having an effect of altering enzyme cleavage activity compared to the unmodified enzyme. Activities associated with the modified DNA cleaving enzyme that can be modulated in different reaction conditions include at least one of: (a) non-sequence specific nicking activity; (b) cleaving the second strand of a duplex DNA at a preexisting nick site to produce a linear duplex with a single strand overhang; (c) non-sequence specific DNA cleavage; (d) cleaving DNA flanking a mismatch; and (e) cleavage at a cruciform structure in a DNA duplex.
摘要翻译：提供了与T7 Endo I具有至少35％的氨基酸序列同一性的修饰的DNA切割酶的组合物和方法。修饰的酶包括两个由“桥”分开的催化中心，其中“桥”至少包含一个突变与未修饰的酶相比具有改变酶裂解活性的作用。与可以在不同反应条件下调节的修饰的DNA切割酶相关的活性包括以下至少一个：（a）非序列特异性切口活性; （b）在预先存在的切口位点切割双链体DNA的第二链以产生具有单链突出端的线性双链体; （c）非序列特异性DNA切割; （d）切断位于不对称侧翼的DNA; 和（e）在DNA双链体中的十字形结构处的切割。

5. 发明申请

US20070042379A1 Modified dna cleavage enzymes and methods for use (as amended by isa) 有权
标题翻译：修饰的dna切割酶和使用方法（由isa修改）
公开(公告)号：US20070042379A1
公开(公告)日：2007-02-22
申请号：US10585964
申请日：2004-11-22
申请人： Chudi Guan , Sanjay Kumar , Rebecca Kucera
发明人： Chudi Guan , Sanjay Kumar , Rebecca Kucera
IPC分类号： C12Q1/68 , C07H21/04 , C12P21/06 , C12N9/22
CPC分类号： C12N9/22
摘要： Compositions and methods are provided that relate to a modified DNA cleaving enzyme having at least 35% amino acid sequence identity with T7 Endo I. The modified enzyme includes two catalytic centers separated by a β-bridge where the β-bridge contains at least one mutation having an effect of altering enzyme cleavage activity compared to the unmodified enzyme. Activities associated with the modified DNA cleaving enzyme that can be modulated in different reaction conditions include at least one of: (a) non-sequence specific nicking activity; (b) cleaving the second strand of a duplex DNA at a preexisting nick site to produce a linear duplex with a single strand overhang; (c) non-sequence specific DNA cleavage; (d) cleaving DNA flanking a mismatch; and (e) cleavage at a cruciform structure in a DNA duplex.
摘要翻译：提供了与T7 Endo I具有至少35％氨基酸序列同一性的修饰的DNA切割酶的组合物和方法。修饰的酶包括由β桥分开的两个催化中心，其中β-桥含有至少一个突变与未修饰的酶相比具有改变酶裂解活性的作用。与可以在不同反应条件下调节的修饰的DNA切割酶相关的活性包括以下至少一个：（a）非序列特异性切口活性; （b）在预先存在的切口位点切割双链体DNA的第二链以产生具有单链突出端的线性双链体; （c）非序列特异性DNA切割; （d）切断位于不对称侧翼的DNA; 和（e）在DNA双链体中的十字形结构处的切割。

6. 发明申请

US20060088868A1 Repair of nucleic acids for improved amplification 有权
标题翻译：修复核酸以改善扩增
公开(公告)号：US20060088868A1
公开(公告)日：2006-04-27
申请号：US11255290
申请日：2005-10-20
申请人： Thomas Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan
发明人： Thomas Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/686 , C12Q1/6844 , C12Q1/6848 , C12Q2521/501 , C12Q2521/301 , C12Q2521/514 , C12Q2521/101 , C12Q2521/331
摘要： Methods and compositions are provided for repairing a polynucleotide so that it can be synthesized efficiently with improved fidelity and yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI. The reaction mixture may further contain an AP endonuclease and a polymerase. These enzymes are optionally selected according to their ability to withstand high temperatures so they can be included in an amplification mixture. The reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture.
摘要翻译：提供了用于修复多核苷酸的方法和组合物，使得其可以在例如扩增反应中以提高的保真度和产率被有效地合成。这包括使用包含连接酶和选自NAD +或ATP的辅因子并在不存在核酸内切酶VI的情况下将多核苷酸与反应混合物一起温育的反应混合物。反应混合物还可以含有AP内切核酸酶和聚合酶。这些酶根据其耐受高温的能力任选地进行选择，因此它们可以被包括在扩增混合物中。反应混合物可以在多核苷酸合成反应之前使用，在这种情况下可以使用不是嗜热的酶。相对于在酶混合物中孵育的秒，分钟或数小时，修复反应不是时间敏感的。

7. 发明授权

US08703462B2 Generation of random double-strand breaks in DNA using enzymes 有权
标题翻译：使用酶产生DNA中的随机双链断裂
公开(公告)号：US08703462B2
公开(公告)日：2014-04-22
申请号：US13140901
申请日：2010-02-03
申请人： Pei-Chung Hsieh , Chudi Guan
发明人： Pei-Chung Hsieh , Chudi Guan
IPC分类号： C12N9/16 , C12N15/00 , C12Q1/68 , C12Q1/34 , C12P19/30 , C07H21/04 , C07H21/02
CPC分类号： C12Q1/6869 , C12Q2525/191 , C12Q2521/301
摘要： An enzyme preparation is described that includes a non-specific nuclease and a T7 Endo I mutant in a unit ratio of less than 1:200. This enzyme preparation may be used to generate double-stranded DNA fragments of a size suitable for DNA sequencing. The ends of the fragments can be readily modified as necessary to ligate adaptors or individual nucleotides to one strand of the double-stranded DNA fragments.
摘要翻译：描述了包含非特异性核酸酶和单体比小于1:200的T7 Endo I突变体的酶制剂。该酶制剂可用于产生适于DNA测序的尺寸的双链DNA片段。可以根据需要容易地修饰片段的末端，以将衔接子或单个核苷酸连接到双链DNA片段的一条链上。

8. 发明授权

US07094563B2 Isolation and composition of novel glycosidases 有权
公开(公告)号：US07094563B2
公开(公告)日：2006-08-22
申请号：US10003136
申请日：2001-11-15
申请人： Sharon T. Wong-Madden , Ellen P. Guthrie , David Landry , Christopher H. Taron , Chudi Guan , Phillips W. Robbins
发明人： Sharon T. Wong-Madden , Ellen P. Guthrie , David Landry , Christopher H. Taron , Chudi Guan , Phillips W. Robbins
IPC分类号： C12P1/00 , C12P19/22 , C12N9/26 , C12N1/00 , C12N1/12
CPC分类号： C12Y302/01051 , C12N9/2402 , C12Y302/0105 , Y10S435/91
摘要： Substantially pure glycosidases capable for cleaving selected glycosidic bonds have been described including glycosidases isolated from Xanthomonas and recombinant glycosidases. Substrate specificity of isolated enzymes have been identified for GlcNacβ1-x, Galα1-3R, Galα1-6R, Galβ1-3R, Fucα-2R, Fucα1-3R, Fucα1-4R, Manα1-2R, Manα1-3R, Manα1-6R, Manβ1-4R, Xylβ1-2R, Glcβ1-4R, and Galβ1-4R providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.

9. 发明授权

US06342365B1 Isolation and composition of novel glycosidases 失效
标题翻译：新型糖苷酶的分离和组成
公开(公告)号：US06342365B1
公开(公告)日：2002-01-29
申请号：US09257153
申请日：1999-02-24
申请人： Sharon T. Wong-Madden , Ellen P. Guthrie , David Landry , Christopher H. Taron , Chudi Guan , Phillips W. Robbins
发明人： Sharon T. Wong-Madden , Ellen P. Guthrie , David Landry , Christopher H. Taron , Chudi Guan , Phillips W. Robbins
IPC分类号： C12Q134
CPC分类号： C12Y302/01051 , C12N9/2402 , C12Y302/0105 , Y10S435/91
摘要： Substantially pure glycosidases capable for cleaving selected glycosidic bonds have been described including glycosidases isolated from Xanthomonas and recombinant glycosidases. Substrate specificity of isolated enzymes have been identified for GlcNac&bgr;1-X, Gal&agr;1-3R, Gal&agr;1-6R, Gal&bgr;1-3R, Fuc&agr;1-2R, Fuc&agr;1-3R, Fuc&agr;1-4R, Man&agr;1-2R, Man&agr;1-3R, Man&agr;1-6R, Man&bgr;1-4R, Xyl&bgr;1-2R, Glc&bgr;1-4R, and Gal&bgr;1-4R providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.
摘要翻译：已经描述了能够切割选择的糖苷键的基本上纯的糖苷酶，其包括从黄单胞菌属和重组糖苷酶分离的糖苷酶。已经鉴定了GlcNacbeta1-X，Galalpha1-3R，Galalpha1-6R，Galbeta1-3R，Fucalpha1-2R，Fucalpha1-3R，Fucalpha1-4R，Manalpha1-2R，Manalpha1-3R，Manalpha1-6R，Manbeta1的分离的酶的底物特异性 -4R，Xylbeta1-2R，Glcbeta1-4R和Galbeta1-4R，其提供了选择性地切割碳水化合物底物中的糖苷键并形成修饰的碳水化合物的改善的能力。

10. 发明授权

US5770405A Isolation and composition of novel glycosidases 失效
标题翻译：新型糖苷酶的分离和组成
公开(公告)号：US5770405A
公开(公告)日：1998-06-23
申请号：US596250
申请日：1996-06-24
申请人： Sharon T. Wong-Madden , Ellen P. Guthrie , Christopher H. Taron , David Landry , Chudi Guan , Phillips W. Robbins
发明人： Sharon T. Wong-Madden , Ellen P. Guthrie , Christopher H. Taron , David Landry , Chudi Guan , Phillips W. Robbins
IPC分类号： C12N15/09 , C12N1/21 , C12N9/24 , C12Q1/34 , C12R1/19 , C12R1/64 , C12P19/44 , C12N1/00 , C12N9/40
CPC分类号： C12Y302/0105 , C12N9/2402 , Y10S435/91
摘要： Purified N-acetylglucosaminidase and .alpha.1-3,6 Galactosidase endogenous to Xanthomonas have been described. Substrate specificity of isolated enzymes have been identified from GlcNAc.beta.1-x and Gal.alpha.1-3R, Gal.alpha.1-6R, providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.
摘要翻译： PCT No.PCT / US94 / 10758 Sec。 371日期：1996年6月5日 102（e）日期1996年6月5日PCT 1994年9月22日PCT公布。出版物WO95 / 08645 日期1995年3月30日已经描述了纯化的N-乙酰氨基葡萄糖苷酶和α1-3,6半乳糖苷酶内源性黄单胞菌属。已经从GlcNAcβ1-x和Galα1-3R，Galα1-6R鉴定了分离的酶的底物特异性，提供了在碳水化合物底物中选择性切割糖苷键并形成改性碳水化合物的改进能力。

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