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1. 发明申请

US20080268507A1 Recombinant Dna Nicking Endonuclease and Uses Thereof 审中-公开
标题翻译：重组Dna Nicking内切核酸酶及其用途
公开(公告)号：US20080268507A1
公开(公告)日：2008-10-30
申请号：US11666148
申请日：2005-10-21
申请人： Shuang-yong Xu , Zhenyu Zhu , Shi-hong Chan , Yan Xu
发明人： Shuang-yong Xu , Zhenyu Zhu , Shi-hong Chan , Yan Xu
IPC分类号： C12P19/34 , C07H21/04 , C12N9/16 , C12N9/10 , C12N15/00 , C12N1/21
CPC分类号： C12N9/22 , C12N9/1241
摘要： Recombinant nicking endonucleases and associated methylases have been obtained and sequenced and their specificity has been defined. A mutant form of the nicking endonuclease has been cloned where the mutation includes deletion of amino acid sequences at the C-terminal end of the protein. The nicking enzymes have been used for a number of purposes including: amplifying DNA from as few cells as can be found in a single bacterial colony in the presence of a strand displacing polymerase; and for removing genomic DNA in a biological preparation where it is deemed to be a contaminant.
摘要翻译：已经获得重组切口内切核酸酶和相关的甲基化酶并测序并确定了它们的特异性。已经克隆了缺口内切核酸酶的突变形式，其中突变包括在蛋白质C末端缺失氨基酸序列。切口酶已经被用于许多目的，包括：在链置换聚合酶存在下，从单个细菌菌落中可以发现的细胞中扩增DNA; 并且用于在被认为是污染物的生物制剂中去除基因组DNA。

2. 发明申请

US20080213860A1 Nicking Endonuclease Methods and Compositions 有权
标题翻译：剔除核酸内切酶方法和组成
公开(公告)号：US20080213860A1
公开(公告)日：2008-09-04
申请号：US11631438
申请日：2005-07-22
申请人： Shuang-Yong Xu , Zhenyu Zhu , Timothy Meixsell
发明人： Shuang-Yong Xu , Zhenyu Zhu , Timothy Meixsell
IPC分类号： C12N9/14
CPC分类号： C12N9/22
摘要： A nicking endonuclease is described which has an amino acid sequence with at least 70% identity to SEQ ID NO:6 and comprising a mutation at least one of an arginine or gutamic acid corresponding to position 507 and position 546 respectively in SEQ ID NO:6.
摘要翻译：描述了一种切口内切核酸酶，其具有与SEQ ID NO：6具有至少70％同一性的氨基酸序列，并且包含分别对应于SEQ ID NO：6中的位置507和位置546的精氨酸或谷氨酸中的至少一种的突变。

3. 发明授权

US06723546B2 Method for cloning and expression of BsaI restriction endonuclease and BsaI methylase in E. coli 有权
标题翻译：在大肠杆菌中克隆和表达BsaI限制性内切核酸酶和BsaI甲基化酶的方法
公开(公告)号：US06723546B2
公开(公告)日：2004-04-20
申请号：US10106275
申请日：2002-03-26
申请人： Zhenyu Zhu , Shuang-Yong Xu
发明人： Zhenyu Zhu , Shuang-Yong Xu
IPC分类号： C12N922
CPC分类号： C12N9/1007 , C12N9/22
摘要： The present invention relates to recombinant DNA which encodes the BsaI restriction endonuclease as well as BsaI methylase, expression of BsaI restriction endonuclease and BsaI methylase in E. coli cells containing the recombinant DNA, and purification of BsaI restriction endonuclease to near homogeneity.
摘要翻译：本发明涉及编码BsaI限制性内切核酸酶以及BsaI甲基化酶的重组DNA，BsaI限制性内切核酸酶和BsaI甲基化酶在含有重组DNA的大肠杆菌细胞中的表达，并将BsaI限制性内切核酸酶纯化至接近同质性。

4. 发明授权

US08361773B2 Polynucleotides encoding polypeptides and host cells therefor 失效
标题翻译：编码多肽的多核苷酸及其宿主细胞
公开(公告)号：US08361773B2
公开(公告)日：2013-01-29
申请号：US12297097
申请日：2007-03-30
申请人： Zhenyu Zhu , Jack S. Benner, II , Shuang-Yong Xu
发明人： Zhenyu Zhu , Jack S. Benner, II , Shuang-Yong Xu
IPC分类号： C07H21/00 , C12P21/00 , C12N1/21 , C12N5/10 , C12N15/00 , C12N9/16 , C07K14/00
CPC分类号： C12N9/22
摘要： Recombinant DNA encoding NruI- and SboI-like restriction endonucleases and methylases and their amino acid sequences are provided as well as methods for expressing the enzymes in transformed host cells and purifying the enzymes.
摘要翻译：提供编码NruI-和SboⅠ样限制性内切核酸酶和甲基化酶及其氨基酸序列的重组DNA以及在转化的宿主细胞中表达酶并纯化酶的方法。

5. 发明授权

US07820424B2 Nicking endonuclease methods and compositions 有权
标题翻译：剔除内切核酸酶方法和组成
公开(公告)号：US07820424B2
公开(公告)日：2010-10-26
申请号：US11631438
申请日：2005-07-22
申请人： Shuang-Yong Xu , Zhenyu Zhu , Timothy Meixsell
发明人： Shuang-Yong Xu , Zhenyu Zhu , Timothy Meixsell
IPC分类号： C12N9/16 , C07K14/00 , C12P21/00 , C12N15/00 , C12N1/21 , C12N5/10 , C12Q1/44 , C07H21/00
CPC分类号： C12N9/22
摘要： A nicking endonuclease is described which has an amino acid sequence with at least 70% identity to SEQ ID NO:6 and comprising a mutation at least one of an arginine or glutamic acid corresponding to position 507 and position 546 respectively in SEQ ID NO:6.
摘要翻译：描述了一种切口内切核酸酶，其具有与SEQ ID NO：6具有至少70％同一性的氨基酸序列，并且包含分别对应于SEQ ID NO：6中的位置507和位置546的精氨酸或谷氨酸中的至少一种的突变。

6. 发明申请

US20090280528A1 Method for Cloning and Expression of NruI Restriction Endonuclease 失效
标题翻译： NruI限制性内切核酸酶的克隆和表达方法
公开(公告)号：US20090280528A1
公开(公告)日：2009-11-12
申请号：US12297097
申请日：2007-03-30
申请人： Zhenyu Zhu , Jack S. Benner, II , Shuang-Yong Xu
发明人： Zhenyu Zhu , Jack S. Benner, II , Shuang-Yong Xu
IPC分类号： C12P21/00 , C07H21/04 , C12N15/63 , C12N1/21 , C12N9/16 , C12N9/10
CPC分类号： C12N9/22
摘要： Recombinant DNA encoding NruI- and SboI-like restriction endonucleases and methylases and their amino acid sequences are provided as well as methods for expressing the enzymes in transformed host cells and purifying the enzymes.
摘要翻译：提供编码NruI-和SboⅠ样限制性内切核酸酶和甲基化酶及其氨基酸序列的重组DNA以及在转化的宿主细胞中表达酶并纯化酶的方法。

7. 发明授权

US06919194B2 Method for cloning and expression of Tth111II restriction endonuclease-methylase in E. coli 失效
标题翻译：在大肠杆菌中克隆和表达Tth111II限制性内切核酸酶 - 甲基化酶的方法
公开(公告)号：US06919194B2
公开(公告)日：2005-07-19
申请号：US10338731
申请日：2003-01-08
申请人： Zhenyu Zhu , Derek Robinson , Jack Benner , Shuang-yong Xu
发明人： Zhenyu Zhu , Derek Robinson , Jack Benner , Shuang-yong Xu
IPC分类号： C12N9/10 , C12N9/22 , C12N15/55
CPC分类号： C12N9/22 , C07K2319/00 , C12N9/1007
摘要： The present invention relates to recombinant DNA which encodes the Tth111II restriction endonuclease-methylase fusion protein (Tth111IIRM), expression of Tth111II restriction endonuclease-methylase fusion protein in E. coli cells containing the recombinant DNA, and purification of Tth111II endonuclease-methylase fusion protein to near homogeneity.
摘要翻译：本发明涉及编码Tth111II限制性内切核酸酶 - 甲基化酶融合蛋白（Tth111IIRM）的重组DNA，含有重组DNA的大肠杆菌细胞中Tth111II限制性内切核酸酶 - 甲基化酶融合蛋白的表达，以及Tth111II内切核酸酶 - 甲基化酶融合蛋白的纯化接近同质性。

8. 发明授权

US06514737B1 Method for cloning and expression of AsiSI restriction endonuclease and AsiSI methylase in E. coli 有权
标题翻译：在大肠杆菌中克隆和表达AsiSI限制性内切核酸酶和AsiSI甲基化酶的方法
公开(公告)号：US06514737B1
公开(公告)日：2003-02-04
申请号：US09933313
申请日：2001-08-20
申请人： Zhenyu Zhu , Shuang-yong Xu
发明人： Zhenyu Zhu , Shuang-yong Xu
IPC分类号： C12N912
CPC分类号： C12N9/1007 , C12N9/22
摘要： The present invention relates to recombinant DNA which encodes the AsiSI restriction endonuclease as well as AsiSI methylase, expression of AsiSI restriction endonuclease and AsiSI methylase in E. coli cells containing the recombinant DNA.
摘要翻译：本发明涉及编码AsiSI限制性内切核酸酶以及AsiSI甲基化酶，AsiSI限制性内切核酸酶和AsiSI甲基化酶在含有重组DNA的大肠杆菌细胞中的表达的重组DNA。

9. 发明授权

US06413758B1 Method for cloning and expression of Bpml restriction endonuclease in E. coli 有权
标题翻译：在大肠杆菌中克隆和表达Bpml限制性内切核酸酶的方法
公开(公告)号：US06413758B1
公开(公告)日：2002-07-02
申请号：US09693146
申请日：2000-10-20
申请人： Shuang-yong Xu , Jian-ping Xiao , Zhenyu Zhu
发明人： Shuang-yong Xu , Jian-ping Xiao , Zhenyu Zhu
IPC分类号： C12N922
CPC分类号： C12N9/22 , C07K2319/00 , C12N9/1007
摘要： The present invention relates to recombinant DNA which encodes the BpmI restriction endonuclease as well as BpmI methyltransferase, expression of BpmI restriction endonuclease from E. coli cells containing the recombinant DNA. BpmI endonuclease is a fusion of two distinct elements with a possible structural domains of restriction-methylation-specificity (R-M-S). This domain organization is analogous to the type I restriction-modification system with three distinct subunits, restriction, methylation, and specificity (R, M, and S). Because BpmI is quite distinct to other type IIs restriction enzymes, it is proposed that BpmI belongs to a subgroup of type II restriction enzymes called type IIf (f stands for fusion of restriction-modification-specificity domains). The Type IIf group of restriction enzyme includes Eco57I, BpmI, GsuI, BseRI and some other restriction enzymes that cut downstream sequences at long distance, 10-20 bp downstream of recognition sequence, such as MmeI (N20/N18)).
摘要翻译：本发明涉及编码BpmI限制性内切核酸酶以及BpmI甲基转移酶的重组DNA，含有重组DNA的大肠杆菌细胞中BpmI限制性内切核酸酶的表达。 BpmI内切核酸酶是具有限制性甲基化特异性（R-M-S）的可能结构域的两个不同元件的融合物。这种结构域组织类似于具有三个不同亚基，限制性，甲基化和特异性（R，M和S）的I型限制性修饰系统。由于BpmI与其他II型限制性内切酶相当不同，所以提出BpmI属于称为IIf型的II型限制酶亚型（f代表限制性修饰特异性结构域的融合）。 IIf类限制酶包括Eco57I，BpmI，GsuI，BseRI和一些其他限制酶，其在识别序列下游10-20bp处长距离切割下游序列，例如MmeI（N20 / N18））。

10. 发明授权

US08372619B2 High fidelity restriction endonucleases 有权
标题翻译：高保真限制性内切核酸酶
公开(公告)号：US08372619B2
公开(公告)日：2013-02-12
申请号：US12172963
申请日：2008-07-14
申请人： Zhenyu Zhu , Aine Blanchard , Shuang-Yong Xu , Shengxi Guan , Hua Wei , Penghua Zhang , Dapeng Sun , Siu-hong Chan
发明人： Zhenyu Zhu , Aine Blanchard , Shuang-Yong Xu , Shengxi Guan , Hua Wei , Penghua Zhang , Dapeng Sun , Siu-hong Chan
IPC分类号： C12N9/22
CPC分类号： C12N15/1058 , C12N9/16 , C12N9/22 , C12N15/1034 , C12N15/1086
摘要： Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions include restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor.
摘要翻译：提供了具有改变性质的酶的组合物和方法，其涉及诱变的系统方法和允许选择所需蛋白质的筛选试验。该方法的实施方案特别适用于修饰限制性核酸内切酶如星形活性的特定性质。组合物包括具有降低的星形活性的限制性内切核酸酶，其由整体保真度指数改善因子定义。

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