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    Artificial promoter libraries for selected organisms and promoters derived from such libraries
    4.
    发明授权
    Artificial promoter libraries for selected organisms and promoters derived from such libraries 失效
    用于所选生物的人工启动子文库和源自这些文库的启动子

    公开(公告)号：US07199233B1

    公开(公告)日：2007-04-03

    申请号：US09242657

    申请日：1997-08-25

    申请人： Peter Ruhdal Jensen ,  Karin Hammer

    发明人： Peter Ruhdal Jensen ,  Karin Hammer

    IPC分类号： C07H21/04 ,  C12Q1/68

    CPC分类号： C12N15/63 ,  C12N15/1051

    摘要： An artificial promoter library (or a set of promoter sequences) for a selected organism or group of organisms is constructed as a mixture of double stranded DNA fragments, the sense strands of which comprise at least two consensus sequences of efficient promoters from said organism or group of organisms, or parts thereof comprising at least half of each, and surrounding intermediate nucleotide sequences (spacers) of variable length in which at least 7 nucleotides are selected randomly among the nucleobases A, T, C and G. The sense strands of the double stranded DNA fragments may also include a regulatory DNA sequence imparting a specific regulatory feature, such as activation by a change in the growth conditions, to the promoters of the library. Further, they may have a sequence comprising one or more recognition sites for restriction endonucleases added to one or both of their ends. The selected organism or group of organisms may be selected from prokaryotes and from eukaryotes; and in prokaryotes the consensus sequences to be retained most often will comprise the −35 signal (−35 to −30): TTGACA and the −10 signal (−12 to −7): TATAAT or parts of both comprising at least 3 conserved nucleotides of each, while in eukaryotes said consensus sequences should comprise a TATA box and at least one upstream activation sequence (UAS). Such artificial promoter libraries can be used, e.g., for optimizing the expression of specific genes in various selected organisms.

    摘要翻译： 构建用于选择的生物体或生物体组的人造启动子文库（或一组启动子序列）作为双链DNA片段的混合物，其有义链包含来自所述生物体或组的至少两个有效启动子的共有序列 的生物体或其部分，其包含每个的至少一半，以及其中在碱基A，T，C和G中随机选择至少7个核苷酸的可变长度的周围中间核苷酸序列（间隔区）。双重的有义链 双链DNA片段还可以包括赋予特异性调节特征的调节性DNA序列，例如通过生长条件的改变来激活文库的启动子。 此外，它们可以具有包含一个或多个加入其一个或两个末端的限制性内切核酸酶识别位点的序列。 选择的有机体或一组生物体可以选自原核生物和真核生物; 并且在原核生物中，最常保留的共有序列将包含-35信号（-35至-30）：TTGACA和-10信号（-12至-7）：TATAAT或两者的部分包含至少3个保守核苷酸 而在真核生物中，共有序列应包含TATA盒和至少一个上游激活序列（UAS）。 可以使用这样的人工启动子文库，例如优化各种选择的生物体中特定基因的表达。