Recombinant Dna Nicking Endonuclease and Uses Thereof
    1.
    发明申请
    Recombinant Dna Nicking Endonuclease and Uses Thereof 审中-公开
    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

    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。

    Nicking Endonuclease Methods and Compositions
    2.
    发明申请
    Nicking Endonuclease Methods and Compositions 有权
    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

    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 。

    Method for cloning and expression of BsaI restriction endonuclease and BsaI methylase in E. coli
    3.
    发明授权
    Method for cloning and expression of BsaI restriction endonuclease and BsaI methylase in E. coli 有权
    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

    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。

    Method for cloning and expression of AsiSI restriction endonuclease and AsiSI methylase in E. coli
    8.
    发明授权
    Method for cloning and expression of AsiSI restriction endonuclease and AsiSI methylase in E. coli 有权
    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

    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。

    Method for cloning and expression of Bpml restriction endonuclease in E. coli
    9.
    发明授权
    Method for cloning and expression of Bpml restriction endonuclease in E. coli 有权
    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

    IPC分类号: C12N922

    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))。