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21 条记录 (耗时 0.029 秒)

    Helicase-dependent amplification of RNA
    12.
    发明授权
    Helicase-dependent amplification of RNA 有权
    RNA的螺旋酶依赖性扩增

    公开(公告)号：US07662594B2

    公开(公告)日：2010-02-16

    申请号：US11327175

    申请日：2006-01-05

    申请人： Huimin Kong ,  Myriam Vincent ,  Yan Xu

    发明人： Huimin Kong ,  Myriam Vincent ,  Yan Xu

    IPC分类号： C12P19/34 ,  C07H21/04 ,  C12N9/00

    CPC分类号： C12P19/34 ,  C12N15/1096

    摘要： Methods and a kit are provided for selectively and exponentially amplifying nucleic acids and include the use of a single strand helicase preparation or a thermostable helicase in the absence of a single strand binding protein and a DNA polymerase such that the amplification can be performed isothermally.

    摘要翻译： 提供了用于选择性和指数扩增核酸的方法和试剂盒，并且包括在不存在单链结合蛋白和DNA聚合酶的情况下使用单链解旋酶制剂或热稳定解旋酶，使得可以等温进行扩增。

    Engineering nicking endonucleases from type IIs restriction endonucleases
    15.
    发明授权
    Engineering nicking endonucleases from type IIs restriction endonucleases 有权
    来自II型限制性内切核酸酶的工程化切口内切核酸酶

    公开(公告)号：US06395523B1

    公开(公告)日：2002-05-28

    申请号：US09872861

    申请日：2001-06-01

    申请人： Huimin Kong ,  Caroline Besnier ,  Yan Xu

    发明人： Huimin Kong ,  Caroline Besnier ,  Yan Xu

    IPC分类号： C12N900

    CPC分类号： C12N9/22

    摘要： The present invention relates to methods to engineer nicking endonucleases from existing Type IIs restriction endonucleases, and the production of the engineered nicking endonucleases. Two engineering methods are disclosed. One involves inactivating the dimerization function of a Type IIs restriction enzyme using site-directed mutagenesis approach. The other involves replacing the cleavage domain of a Type IIs restriction enzyme with the cleavage domain from a natural occurring nicking endonuclease, N.BstNBI.

    摘要翻译： 本发明涉及从现有的II型限制性内切核酸酶工程化切口内切核酸酶的方法，以及工程化切口内切核酸酶的产生。 公开了两种工程方法。 一个涉及使用定点诱变方法使II型限制酶的二聚功能失活。 另一个涉及用来自天然存在的切口内切核酸酶N.BstNBI的切割结构域替换II型限制酶的切割结构域。

    Method for cloning and producing the SnaBI restriction endonuclease and purification of the recombinant SnaBI restriction endonuclease
    16.
    发明授权
    Method for cloning and producing the SnaBI restriction endonuclease and purification of the recombinant SnaBI restriction endonuclease 有权
    用于克隆和产生SnaBI限制性内切核酸酶并纯化重组SnaBI限制性内切核酸酶的方法

    公开(公告)号：US6025179A

    公开(公告)日：2000-02-15

    申请号：US143776

    申请日：1998-08-31

    申请人： Keith D. Lunnen ,  Huimin Kong ,  Geoffrey G Wilson

    发明人： Keith D. Lunnen ,  Huimin Kong ,  Geoffrey G Wilson

    IPC分类号： C12N15/09 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N9/10 ,  C12N9/16 ,  C12N9/22 ,  C12N15/52 ,  C12N15/55

    CPC分类号： C12N15/52 ,  C12N9/1007 ,  C12N9/22

    摘要： The methylase selection method was used to clone the SnaBI methylase gene (snaBIM) from Sphaerotilus natans (ATCC 15291). An active SnaBI methylase was cloned in E. coli using pSnaBI-2, a pUC19 derivative containing two SnaBI sites. Because methylase and restriction genes are usually located alongside each other in a restriction-modification systems, efforts were made to clone the downstream DNA by inverse PCR. Inverse PCR cloned the missing portion of the SnaBI endonuclease and also identified a control, or C, protein. The two open reading frames snaBIR (ORF1) and snaBIC (ORF2) converged toward the SnaBI methylase gene (ORF). Attempts to establish a snaBIR-recombinant plasmid expressing the SnaBI endonuclease in E.coli modified with SnaBI methylase failed. Overexpression of the SnaBI endonuclease in E. coli required the use of the heterospecific BsaAI methylase.

    摘要翻译： 甲基化酶选择方法用于从Sphaerotilus natans（ATCC 15291）克隆SnaBI甲基化酶基因（snaBIM）。 使用含有两个SnaBI位点的pUC19衍生物pSnaBI-2将活性SnaBI甲基化酶克隆到大肠杆菌中。 因为甲基化酶和限制性基因通常在限制性修饰系统中彼此并列，所以努力通过反向PCR克隆下游DNA。 反向PCR克隆了SnaBI内切核酸酶的缺失部分，并鉴定了对照或C蛋白。 snaBIR（ORF1）和snaBIC（ORF2）两个开放阅读框向SnaBI甲基化酶基因（ORF）收敛。 在SnaBI甲基化酶修饰的大肠杆菌中建立表达SnaBI内切核酸酶的snaBIR重组质粒的尝试失败。 大肠杆菌中SnaBI内切核酸酶的过表达需要使用异源特异性BsaAI甲基化酶。

    Helicase-dependent amplification of circular nucleic acids
    19.
    发明申请
    Helicase-dependent amplification of circular nucleic acids 审中-公开
    循环核酸的螺旋酶依赖性扩增

    公开(公告)号：US20100075384A1

    公开(公告)日：2010-03-25

    申请号：US10594095

    申请日：2005-03-23

    申请人： Huimin Kong ,  Yan Xu

    发明人： Huimin Kong ,  Yan Xu

    IPC分类号： C12P19/34 ,  C12N9/10

    CPC分类号： C12P19/34 ,  C12Q1/6844 ,  C12Q2531/125 ,  C12Q2521/513

    摘要： A helicase-mediated amplification method for circular DNA templates and target DNA sequences within the templates is provided. The method combines a DNA polymerase and a helicase preparation to amplify a target sequence as well as the entire circular DNA template.

    摘要翻译： 提供了解旋酶介导的扩增方法，用于模板内的循环DNA模板和靶DNA序列。 该方法结合DNA聚合酶和解旋酶制剂以扩增靶序列以及整个环状DNA模板。