Hyperactive transposase mutants of the Himar1 mariner transposon
Hyperactive transposase mutants of the Himar1 mariner transposon
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DOI：
10.1073/pnas.96.20.11428
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发表时间：
1999-09-28
影响因子：
11.1
通讯作者：
Robertson, HM
中科院分区：
文献类型：
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作者：
Lampe, DJ;Akerley, BJ;Robertson, HM
Mariner 家族转座元件在多种生物体中都很活跃，并且在缺乏复杂遗传学的物种中正成为越来越重要的遗传工具。从角蝇、Haematobia irritans 中分离出来的 Himar1 元件在适当表达时在大肠杆菌中也很活跃。我们利用这一事实设计了一种针对 Hinnar1 转座酶高活性突变体的遗传筛选，根据大肠杆菌测定的结果，该突变体将整体转座增强了约 4 至 50 倍。在体外转座测定中，纯化的突变转座酶保留了其高活性，尽管程度较低。像本文描述的突变体应该能够对水手转座的生物化学进行复杂的分析，并且应该改进这些元件作为遗传工具的体内和体外的使用。
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, isolated from the horn fly, Haematobia irritans, is active in Escherichia coli when expressed appropriately. We used this fact to devise a genetic screen for hyperactive mutants of Hinnar1 transposase that enhance overall transposition from approximate to 4- to 50-fold as measured in an E. coli assay. Purified mutant transposases retain their hyperactivity, although to a lesser degree, in an in vitro transposition assay. Mutants like those described herein should enable sophisticated analysis of the biochemistry of mariner transposition and should improve the use of these elements as genetic tools, both in vivo and in vitro.