【2h】

Hyperactive transposase mutants of the Himar1 mariner transposon

机译:Himar1 Mariner转座子的高活性转座酶突变体

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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, 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 Himar1 transposase that enhance overall transposition from ≈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.
机译:水手家族的转座因子在多种生物中都有活性,并且在缺乏复杂遗传学的物种中正变得越来越重要。从角蝇蝇中分离出的Himar1元件在适当表达时对大肠杆菌具有活性。我们利用这一事实为Himar1转座酶的高活性突变体设计了一个基因筛选,该突变体可将整体转座率从≈4倍提高到50倍(如在大肠杆菌测定中所测)。在体外转座试验中,纯化的突变转座酶虽然程度较小,但仍保留其高活性。像本文所述的那些突变体应该能够对水手转座的生物化学进行复杂的分析,并且应该改善体内和体外这些元件作为遗传工具的用途。

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