Hyperactive transposase mutants of the Himar1 mariner transposon

被引:204
作者
Lampe, DJ [1 ]
Akerley, BJ
Rubin, EJ
Mekalanos, JJ
Robertson, HM
机构
[1] Duquesne Univ, Dept Biol Sci, Pittsburgh, PA 15282 USA
[2] Harvard Univ, Sch Med, Dept Microbiol & Mol Genet, Boston, MA 02115 USA
[3] Univ Illinois, Dept Entomol, Urbana, IL 61801 USA
关键词
D O I
10.1073/pnas.96.20.11428
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
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 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.
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页码:11428 / 11433
页数:6
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