Hyperactive mutants of Himar1 transposase and methods for using the same
    3.
    发明授权
    Hyperactive mutants of Himar1 transposase and methods for using the same 失效
    Himar1转座酶的活性突变体及其使用方法

    公开(公告)号:US06368830B1

    公开(公告)日:2002-04-09

    申请号:US09671950

    申请日:2000-09-27

    IPC分类号: C12N1574

    CPC分类号: C12N9/22 C12N15/74

    摘要: Mariner-family transposable elements are active in a wide variety of organisms and are becoming increasingly important genetic tools in species lacking sophisticated genetics. The Himar1 element, a member of the mariner family, isolated from the horn fly, Haematobia irritans, is active in Escherichia coli when expressed appropriately. Using this fact, a genetic screen was devised to isolate hyperactive mutants of Himar1 transposase that enhance overall transposition from 4 to 50-fold as measured in an E. coli assay. These hyperactive Himar1 mutant transposases should enable sophisticated analysis of the biochemistry of mariner transposition and should improve efficiency of a variety of genetic manipulations involving transposition in vivo and in vitro such as random mutagenesis or transgenesis in a wide range of host cells than the transposable elements previously available.

    摘要翻译: 水手家族转座因子在各种生物体中是活跃的,并且在缺乏复杂遗传学的物种中变得越来越重要的遗传工具。 Himar1元素,海员家族的成员,从角飞,哈马氏杆菌分离,在适当表达时在大肠杆菌中活跃。 使用这个事实,设计了一种遗传筛选来分离Himar1转座酶的活性突变体,其将整体转座增加4倍至50倍,如在大肠杆菌测定中所测定的。 这些超活跃的Himar1突变体转座酶应该能够复杂地分析海员转座的生物化学,并且应该提高涉及体内和体外转座的各种遗传操作的效率,例如在广泛的宿主细胞中的随机诱变或转基因,而不是以前的转座元件 可用。