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44 条记录 (耗时 0.066 秒)

    Saturation mutagenesis in directed evolution
    21.
    发明申请
    Saturation mutagenesis in directed evolution 审中-公开
    定向进化中的饱和诱变

    公开(公告)号：US20050100985A1

    公开(公告)日：2005-05-12

    申请号：US10644410

    申请日：2003-08-20

    申请人： Jay Short

    发明人： Jay Short

    IPC分类号： A61K39/00 ,  C07K14/445 ,  C12N5/02 ,  C12N9/00 ,  C12N9/16 ,  C12N9/24 ,  C12N9/38 ,  C12N15/10 ,  C12Q1/68 ,  C12P21/02 ,  C07K14/47 ,  C12N5/06 ,  C12P19/34

    CPC分类号： C07K14/445 ,  A61K39/00 ,  A61K2039/53 ,  C12N9/00 ,  C12N9/16 ,  C12N15/102 ,  C12N15/1027 ,  C12N15/1034 ,  C12N15/1058

    摘要： Disclosed is a rapid and facilitated method of producing from a parentlal template polynucleotide, a set of mutagenized progeny polynculeotides whereby at each original codon position there is produced at least one substitute codon encoding each of the 20 naturally encoded amino acids. Accordingly, there is also provided a method of producing from a parental template polypeptide, a set of mutagenized progeny polypeptides wherein each of the 20 naturally encoded amino acids is represented at each original amino acid position. The method provided is termed site-saturation mutagenesis, or simply saturation mutagenesis, and can be used in combination with other mutagenization processes, such as, for example, a process wherein two or more related polynucleotides are introduced into a suitable host cell such that a hybrid polynucleotide is generated by recombination and reductive reassortment. Also provided are vector and expression vehicles incuding such polynucleotides, polypeptides expressed by the hybrid polynucleotides and a method for screening for hybrid polypeptides.

    摘要翻译： 公开了一种由亲本模板多核苷酸，一组诱变的后代多核苷酸产生的快速和便利的方法，其中在每个原始密码子位置产生编码20个天然编码氨基酸中的每一个的至少一个替代密码子。 因此，还提供了从亲本模板多肽，一组诱变的后代多肽生产其中每个原始氨基酸位置上表示20个天然编码氨基酸的方法。 所提供的方法称为位点饱和诱变或简单的饱和诱变，并且可以与其它诱变过程组合使用，例如其中将两个或更多个相关多核苷酸引入合适的宿主细胞中的方法，使得 通过重组和还原重配产生杂交多核苷酸。 还提供了包含这种多核苷酸的载体和表达载体，由杂交多核苷酸表达的多肽和杂交多肽的筛选方法。

    Polypeptides having polymerase activity and methods of use thereof
    25.
    发明授权
    Polypeptides having polymerase activity and methods of use thereof 失效
    具有聚合酶活性的多肽及其使用方法

    公开(公告)号：US07056703B2

    公开(公告)日：2006-06-06

    申请号：US09948369

    申请日：2001-09-06

    申请人： Walter Callen ,  Eric J. Mathur ,  Jay Short

    发明人： Walter Callen ,  Eric J. Mathur ,  Jay Short

    IPC分类号： C12P19/34 ,  C12Q1/68 ,  C12N9/12

    CPC分类号： C12N9/1252 ,  A01K2217/05

    摘要： The invention relates to thermostable polymerases that have polymerase activity temperatures in the range from 90° C. up to 113° C., such as those derived from Pyrolobus fumaria, and to polynucleotides encoding the polymerases In addition, methods of designing new thermostable DNA polymerases and methods of use thereof are also provided. The polymerases have increased activity and stability at increased pH and temperature.

    摘要翻译： 本发明涉及聚合酶活性温度在90℃至113℃范围内的热稳定聚合酶，例如源自富马酸火焰菌的那些，以及编码聚合酶的多核苷酸。此外，设计新的耐热DNA聚合酶的方法 还提供了其使用方法。 聚合酶在增加的pH和温度下具有增加的活性和稳定性。

    High throughput screening for novel bioactivities
    29.
    发明申请
    High throughput screening for novel bioactivities 审中-公开
    高通量筛选新型生物活性

    公开(公告)号：US20050040550A1

    公开(公告)日：2005-02-24

    申请号：US10948807

    申请日：2004-09-23

    申请人： Jay Short ,  Martin Keller

    发明人： Jay Short ,  Martin Keller

    IPC分类号： C12N15/09 ,  C12N15/10 ,  C12Q1/68 ,  C40B40/02 ,  B01J13/02 ,  B01J13/04

    CPC分类号： C40B40/02 ,  C12N15/1037 ,  C12N15/1055 ,  C12Q1/6811

    摘要： Disclosed is a process for identifying clones having a specified activity of interest, which process comprises (i) generating one or more expression libraries derived from nuclei acid directly isolated from the environment; and (ii) screening said libraries utilizing a fluorescence activated cell sorter to identify said clones. More particularly, this is a process for identifying clones having a specified activity of interest by (i) generating one or more expression libraries derived from nucleic acid directly or indirectly isolated from the environment; (ii) exposing said libraries to a particular substrate or substrates of interest; and (iii) screening said exposed libraries utilizing a fluorescence activated cell sorter to identify clones which react with the substrate or substrates. Also provided is a process for identifying clones having a specified activity of interest by (i) generating one or more expression libraries derived from nucleic acid directly or indirectly isolated from the environment; and (ii) screening said exposed libraries utilizing an assay requiring co-encapsulation, a binding event or the covalent modification of a target, and a fluorescence activated cell sorter to identify positive clones.

    摘要翻译： 公开了用于鉴定具有特定的感兴趣活性的克隆的方法，该方法包括（i）产生一种或多种衍生自从环境直接分离的核酸的表达文库; 和（ii）使用荧光激活细胞分选仪筛选所述文库以鉴定所述克隆。 更具体地说，这是通过（i）产生一个或多个从环境中直接或间接分离的核酸产生的表达文库来鉴定具有特定活性的克隆的方法; （ii）将所述文库暴露于感兴趣的特定底物或底物; 和（iii）使用荧光激活的细胞分选仪筛选所述暴露的文库，以鉴定与底物或底物反应的克隆。 还提供了通过（i）生成从环境直接或间接分离的核酸产生的一种或多种表达文库来鉴定具有特定活性的克隆的方法; 和（ii）使用需要共包封，结合事件或靶共价修饰的测定法和荧光激活的细胞分选器筛选所述暴露的文库以鉴定阳性克隆。