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8 条记录 (耗时 0.016 秒)

    MULTIPLE LIGASE COMPOSITIONS, SYSTEMS, AND METHODS
    4.
    发明申请
    MULTIPLE LIGASE COMPOSITIONS, SYSTEMS, AND METHODS 审中-公开

    公开(公告)号：US20170226498A1

    公开(公告)日：2017-08-10

    申请号：US15426543

    申请日：2017-02-07

    申请人： RGENE, Inc.

    发明人： Yu Zheng ,  Manqing Hong

    IPC分类号： C12N9/00

    CPC分类号： C12N9/93 ,  C12Y605/01001 ,  C12Y605/01002

    摘要： Provided herein are compositions, systems, and methods using multiple ligases, wherein at least one of the ligases is an adenylation-deficient ATP-dependent ligase or an un-adenylated ATP-dependent ligase (e.g., present in an ATP free mixture). In certain embodiments, multiple ligases are used to ligate a pre-adenylated double stranded sequence to a non-adenylated double stranded sequence (e.g., the adenylation-deficient ATP-dependent ligase or un-adenylated ATP-dependent ligase ligates the first strand, and a second ligase ligates the second strand). In other embodiments, provided herein are mutant T4 ligases (e.g., K159S mutant or K159C mutant).

    BIOSENSORS THAT DETECT NAD+
    5.
    发明申请
    BIOSENSORS THAT DETECT NAD+ 审中-公开
    检测NAD +的生物传感器

    公开(公告)号：US20160153023A1

    公开(公告)日：2016-06-02

    申请号：US14948161

    申请日：2015-11-20

    申请人： Richard Goodman ,  Michael Cohen ,  Lulu Cambronne ,  Melissa Stewart

    发明人： Richard Goodman ,  Michael Cohen ,  Lulu Cambronne ,  Melissa Stewart

    IPC分类号： C12Q1/25 ,  C12N9/00

    CPC分类号： C12Q1/25 ,  C12N9/93 ,  C12Q1/008 ,  C12Y605/01002 ,  G01N2333/9015

    摘要： A polypeptide biosensor that detects free NAD+ is disclosed. The polypeptide comprises a first fragment from an NAD+ dependent DNA ligase acetylation domain, a second fragment from the NAD+ dependent DNA ligase acetylation domain, and a fluorescent protein, wherein the fluorescent protein is positioned between the two DNA ligase acetylation domain fragments. Also disclosed are expression vectors comprising the biosensor as well as methods of using the biosensor to detect NAD+.

    摘要翻译： 公开了检测游离NAD +的多肽生物传感器。 多肽包含来自NAD +依赖性DNA连接酶乙酰化结构域的第一片段，来自NAD +依赖性DNA连接酶乙酰化结构域的第二片段和荧光蛋白，其中所述荧光蛋白位于两个DNA连接酶乙酰化结构域片段之间。 还公开了包含生物传感器的表达载体以及使用生物传感器检测NAD +的方法。

    Improved Methods for Rapid Gene Synthesis
    8.
    发明申请
    Improved Methods for Rapid Gene Synthesis 审中-公开
    改进的快速基因合成方法

    公开(公告)号：US20120156731A1

    公开(公告)日：2012-06-21

    申请号：US12052156

    申请日：2008-03-20

    申请人： Yanyi Huang ,  James Berger ,  Stephen Quake

    发明人： Yanyi Huang ,  James Berger ,  Stephen Quake

    IPC分类号： C12P19/34 ,  C07H21/04 ,  C12M1/00 ,  C12N9/00

    CPC分类号： C12Y605/01001 ,  C12P19/34 ,  C12Y605/01002

    摘要： Disclosed are methods and materials for assembling long polynucleotides from synthetic oligonucleotides. The use of synthetic oligonucleotides permits non-natural design of sequences. The oligonucleotides used for construction may be relatively short, according to practicalities of nucleotide synthesis. They are assembled using a ligase which is operative over a range of temperatures, i.e., is thermostable. The method and oligonucleotides are designed such that the melting temperature of the strands to be hybridized is set at a number of selected specific temperatures for each group of oligonucleotides to be hybridized and ligated. Hybridization and ligation take place at or near the melting temperature, so that each succeeding ligation is governed by a temperature that will prevent hybridization if any mismatches are present.

    摘要翻译： 公开了用于从合成寡核苷酸组装长多核苷酸的方法和材料。 合成寡核苷酸的使用允许序列的非自然设计。 根据核苷酸合成的实用性，用于构建的寡核苷酸可能相对较短。 它们使用在一定温度范围内操作的连接酶组装，即是耐热的。 设计该方法和寡核苷酸使待杂交的链的解链温度被设定为要混合并连接的每组寡核苷酸的选定的特定温度的数目。 杂交和连接在熔融温度或接近熔化温度下进行，因此每次后续连接都由温度控制，如果存在任何错配，将会阻止杂交。